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Keywords = lipid peroxidation

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26 pages, 1384 KiB  
Article
A Moderate Intake of Beer Improves Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) in a High-Fat Diet (HFD)-Induced Mouse Model
by Andrea Vornoli, Aymen Souid, Barbara Lazzari, Federica Turri, Flavia Pizzi, Emilia Bramanti, Beatrice Campanella, Cheherazade Trouki, Andrea Raffaelli, Marta Wójcik, Clara Maria Della Croce, Lucia Giorgetti, Vincenzo Longo, Emanuele Capra and Luisa Pozzo
Molecules 2024, 29(24), 5954; https://doi.org/10.3390/molecules29245954 - 17 Dec 2024
Abstract
Beer and its components show potential for reducing hepatic steatosis in rodent models through multiple mechanisms. This study aimed to evaluate beer’s anti-steatotic effects in a high-fat diet (HFD)-induced mouse model of Metabolic dysfunction-Associated Liver Disease (MASLD) and to explore the underlying mechanisms. [...] Read more.
Beer and its components show potential for reducing hepatic steatosis in rodent models through multiple mechanisms. This study aimed to evaluate beer’s anti-steatotic effects in a high-fat diet (HFD)-induced mouse model of Metabolic dysfunction-Associated Liver Disease (MASLD) and to explore the underlying mechanisms. In the HFD group, steatosis was confirmed by altered blood parameters, weight gain, elevated liver lipid content, and histological changes. These markers were normalized in the HFD+beer group, reaching levels similar to the control (CTR) group. Protein carbonylation and lipid peroxidation levels were consistent across all groups, suggesting that the model represents an early stage of MASLD without oxidative stress. Transcriptomic and CpG methylation analyses revealed clear distinctions between the CTR and HFD groups. RNA sequencing identified 162 differentially expressed genes (DEGs) between the CTR and HFD groups, primarily related to inflammation and lipid regulation. Beer consumption modified the health of the HFD mice, affecting inflammation but not lipid homeostasis (CTR vs. HFD+beer, DEGs = 43). The CpG methylation analysis indicated that beer lowered methylation, impacting genes linked to lipid accumulation and inflammation. A cecal metabolite analysis suggested that beer improved short-chain fatty acid metabolism (SCFA). In summary, a moderate beer intake may mitigate MASLD by modulating lipid metabolism and SCFA pathways, likely through polyphenol activity. Full article
(This article belongs to the Special Issue Advances in Natural Products and Their Biological Activities)
15 pages, 2787 KiB  
Article
Comparative Salt-Stress Responses in Salt-Tolerant (Vikinga) and Salt-Sensitive (Regalona) Quinoa Varieties. Physiological, Anatomical and Biochemical Perspectives
by Xavier Serrat, Antony Quello, Brigen Manikan, Gladys Lino and Salvador Nogués
Agronomy 2024, 14(12), 3003; https://doi.org/10.3390/agronomy14123003 - 17 Dec 2024
Abstract
Soil salinization is an important stress factor that limits plant growth and yield. Increased salinization is projected to affect more than 50% of all arable land by 2050. In addition, the growing demand for food, together with the increase in the world population, [...] Read more.
Soil salinization is an important stress factor that limits plant growth and yield. Increased salinization is projected to affect more than 50% of all arable land by 2050. In addition, the growing demand for food, together with the increase in the world population, forces the need to seek salt-tolerant crops. Quinoa (Chenopodium quinoa Willd.) is an Andean crop of high importance, due to its nutritional characteristics and high tolerance to different abiotic stresses. The aim of this work is to determine the physiological, anatomical, and biochemical salt-tolerance mechanisms of a salt-tolerant (Vikinga) and a salt-sensitive (Regalona) quinoa variety. Plants were subjected to salinity stress for 15 days, starting at 100 mM NaCl until progressively reaching 400 mM NaCl. Physiological, anatomical, and biochemical parameters including growth, chlorophyll content, quantum yield of PSII (ϕPSII), gas exchange, stomatal density, size, and lipid peroxidation (via malondialdehyde, MDA) were measured. Results show that chlorophyll content, ϕPSII, and MDA were not significantly reduced under saline stress in both varieties. The most stress-affected process was the CO2 net assimilation, with an up to 60% reduction in both varieties, yet Vikinga produced higher dry weight than Regalona due to the number of leaves. The stomatal densities increased under salinity for both varieties, with Regalona the one showing higher values. The averaged stomatal size was also reduced under salinity in both varieties. The capacity of Vikinga to generate higher dry weight is a function of the capacity to generate greater amounts of leaves and roots in any condition. The stomatal control is a key mechanism in quinoa’s salinity tolerance, acquiring higher densities with smaller sizes for efficient management of water loss and carbon assimilation. These findings highlight the potential of Vikinga for cultivation in temperate salinized environments during winter, such as Deltas and lowlands where rice is grown during summer. Full article
(This article belongs to the Section Plant-Crop Biology and Biochemistry)
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Figure 1

Figure 1
<p>Scheme summarizing the experiment. Grey to black colors represent low (100 mM) to high (400 mM) NaCl concentration in the nutritive solutions.</p>
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<p>Growth measurements of Vikinga and Regalona varieties under salinity conditions (400 mM NaCl): (<b>A</b>) plant height (cm); (<b>B</b>) fresh weight of the whole plant (g) and fresh weight of the leaves (dark grey), stem (pale grey), and root (black); (<b>C</b>) shoot/root ratio; (<b>D</b>) dry weight (g) of the whole plant and dry weight of the leaves (dark grey), stem (pale grey), and root (black). Data are means of twenty-four repetitions (<span class="html-italic">n</span> = 24) ± standard error. According to Tukey’s multiple-comparisons test, significant differences with a <span class="html-italic">p</span> &lt; 0.05 have been represented with different letters.</p>
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<p>(<b>A</b>) SPAD, (<b>B</b>) quantum yield of photosystem II, (<b>C</b>) net CO<sub>2</sub> assimilation (A<sub>n</sub>), (<b>D</b>) stomatal conductance (g<sub>s</sub>), (<b>E</b>) intercellular concentration of CO<sub>2</sub> (C<sub>i</sub>), and (<b>F</b>) leaf temperature for Vikinga (closed bars) and Regalona (open bars) varieties. Data are means of twenty-four repetitions (n = 24) ±standard error. According to Tukey’s multiple-comparisons test, significant differences with a <span class="html-italic">p</span> &lt; 0.05 have been represented with different letters.</p>
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<p>(<b>A</b>) Stomatal density of the adaxial surface of the leaves and (<b>B</b>) Stomatal density of the abaxial surface of the leaves for Vikinga (black bars) and Regalona (white bars) varieties. According to Tukey’s multiple—comparisons test, significant differences with a <span class="html-italic">p</span> &lt;0.05.</p>
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<p>(<b>A</b>,<b>B</b>) Microscopic images of the adaxial and abaxial surface of the Vikinga variety in the control treatment. (<b>C</b>,<b>D</b>) Microscopic images of the adaxial and abaxial surface of the Regalona variety under control treatment. (<b>E</b>,<b>F</b>) Microscopic images of the adaxial and abaxial surface, respectively, of the Vikinga variety in the saline treatment (400 mM NaCl). (<b>G</b>,<b>H</b>) Microscopic images of the adaxial and abaxial surface, respectively, of the Regalona variety in the saline treatment (400 mM NaCl).</p>
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<p>Length and width of adaxial and abaxial stomata comparing Vikinga and Regalona under salinity (400 mM NaCl) and control conditions (0 mM NaCl). Data means of thirty-two repetitions (n = 32) ± standard error. According to Tukey’s multiple-comparisons test, significant differences with a <span class="html-italic">p</span> &lt; 0.05 have been represented with capital letters when comparing stomata lengths and lower-case letters when comparing widths.</p>
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<p>MDA values (nm g<sup>−1</sup> PF) for Vikinga (black bars) and Regalona (white bars). The MDA data are means of four repetitions (n = 4). According to Tukey’s multiple-comparisons test, significant differences with a <span class="html-italic">p</span> &lt; 0.05 have been represented with different letters.</p>
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18 pages, 9603 KiB  
Article
High Carbonyl Graphene Oxide Suppresses Colorectal Cancer Cell Proliferation and Migration by Inducing Ferroptosis via the System Xc−/GSH/GPX4 Axis
by Xiecheng Zhou, Qixing Zhang, Haoran Zhu, Guangxiong Ouyang, Xin Wang and Yuankun Cai
Pharmaceutics 2024, 16(12), 1605; https://doi.org/10.3390/pharmaceutics16121605 - 17 Dec 2024
Viewed by 188
Abstract
Background/Objectives: Colorectal cancer (CRC) is characterized by a high rate of both incidence and mortality, and its treatment outcomes are often affected by recurrence and drug resistance. Ferroptosis, an iron-dependent programmed cell death mechanism triggered by lipid peroxidation, has recently gained attention as [...] Read more.
Background/Objectives: Colorectal cancer (CRC) is characterized by a high rate of both incidence and mortality, and its treatment outcomes are often affected by recurrence and drug resistance. Ferroptosis, an iron-dependent programmed cell death mechanism triggered by lipid peroxidation, has recently gained attention as a potential therapeutic target. Graphene oxide (GO), known for its oxygen-containing functional groups, biocompatibility, and potential for functionalization, holds promise in cancer treatment. However, its role in ferroptosis induction in CRC remains underexplored. The objective of this study was to investigate the effects of High Carbonyl Graphene Oxide (HC-GO) on ferroptosis in CRC and elucidate the underlying mechanisms. Methods: In vitro assays were conducted to evaluate the impact of HC-GO on CRC cell proliferation, mitochondrial function, iron accumulation, lipid peroxidation, and reactive oxygen species (ROS) production. The ferroptosis inhibitor Fer-1 was used to confirm the role of ferroptosis in HC-GO’s anti-tumor effects. In vivo, the anti-tumor activity of HC-GO was assessed in a CRC xenograft model, with organ toxicity evaluated. Results: HC-GO significantly inhibited CRC cell proliferation, induced mitochondrial damage, and enhanced iron accumulation, lipid peroxidation, and ROS production. It also downregulated the ferroptosis-inhibiting proteins GPX4 and SLC7A11, which were reversed by Fer-1, confirming the involvement of ferroptosis in HC-GO’s anti-cancer effects. In vivo, HC-GO significantly suppressed tumor growth without noticeable toxicity to vital organs. Conclusions: HC-GO triggered ferroptosis in CRC cells by suppressing the System Xc−/GSH/GPX4 pathway, providing a novel therapeutic strategy for CRC treatment. These findings suggest HC-GO as a promising nanomedicine for clinical application, warranting further investigation to explore its potential in CRC therapy. Full article
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Figure 1
<p>Characterization of HC-GO. (<b>A</b>) Scanning electron microscope images of HC-GO, scale bar: 2 μm, 10 μm; (<b>B</b>) AFM topography image and the corresponding height distribution graph of HC-GO, scale bar: 2 μm; (<b>C</b>) Raman spectra results of HC-GO; (<b>D</b>) XPS results comparing regular GO and HC-GO.</p>
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<p>HC-GO significantly inhibited the in vitro proliferation and migration of HCT116 and HCT15 cells. (<b>A</b>,<b>B</b>) Colony formation assays were used to analyze cell proliferation. (<b>C</b>,<b>D</b>) CCK-8 assays were conducted to assess cell proliferation. (<b>E</b>,<b>F</b>) Scratch wound healing assays were employed to analyze cell migration. (<b>G</b>,<b>H</b>) Transwell assays were performed to measure cell migration capacity. Scale bar: 100 μm. (<b>I</b>,<b>J</b>) Western blot (WB) analysis was used to assess the expression levels of stemness proteins. Data are presented as mean ± SD. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 compared with the control group. All experiments were independently repeated at least three times.</p>
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<p>HC-GO induced ferroptosis in HCT116 and HCT15 cells in vitro. (<b>A</b>) Analysis of Fe<sup>2+</sup> levels. (<b>B</b>,<b>C</b>) Analysis of intracellular ROS levels. (<b>D</b>,<b>E</b>) Analysis of lipid ROS levels. (<b>F</b>) Analysis of GSH levels. (<b>G</b>,<b>H</b>) Western blot analysis of ferroptosis-related protein expression levels. (<b>I</b>) Transmission electron microscopy images of HCT116 cells: blue arrows indicate damaged mitochondria, red arrows indicate mitochondria with vacuolization, and yellow circles highlight structural damage in mitochondria (increased membrane density, reduced cristae, and mitochondrial shrinkage). Scale bars: 1 μm, 2 μm, 500 nm. Data are presented as mean ± SD. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001 compared with the control group. All experiments were independently repeated at least three times.</p>
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<p>HC-GO inhibited CRC cells in vivo. (<b>A</b>,<b>B</b>) Tumor appearance; (<b>C</b>,<b>D</b>) Tumor volume and weight. Data are presented as mean ± SD. ** <span class="html-italic">p</span> &lt; 0.01 compared with the control group.</p>
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<p>HC-GO induced ferroptosis in vivo. (<b>A</b>) HE staining used to assess morphological differences in tumor tissues. Scale bar: 200 μm. (<b>B</b>,<b>C</b>) Immunohistochemical staining for GPX4 and SLC7A11. Scale bar: 200 μm. (<b>D</b>) HE staining used to assess the morphology of mouse heart, liver, spleen, lung, and kidney tissues. Scale bar: 200 μm. Data are presented as mean ± SD. **** <span class="html-italic">p</span> &lt; 0.0001compared with the control group.</p>
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<p>Ferroptosis inhibitor Fer-1 blocked HC-GO-induced ferroptosis in HCT116 cells. The concentration of Fer-1 was 10 μM [<a href="#B39-pharmaceutics-16-01605" class="html-bibr">39</a>]. (<b>A</b>) CCK-8 assay used to analyze cell proliferation; (<b>B</b>,<b>C</b>) Colony formation assay used to analyze cell proliferation; (<b>D</b>,<b>E</b>) Fer-1 blocks the increase in intracellular ROS levels induced by HC-GO; (<b>F</b>,<b>G</b>) Fer-1 blocks the increase in intracellular lipid ROS levels induced by HC-GO; (<b>H</b>) Fer-1 blocks the increase in GSH levels induced by HC-GO; (<b>I</b>,<b>J</b>) Western blot (WB) results show that Fer-1 blocks the decrease in GPX4 and SLC7A11 expression induced by HC-GO; (<b>K</b>,<b>L</b>) Immunofluorescence shows that Fer-1 blocks the decrease in GPX4 expression induced by HC-GO; (<b>M</b>,<b>N</b>) Immunofluorescence shows that Fer-1 blocks the decrease in SLC7A11 expression induced by HC-GO. Scale bar: 50 μm. Data are presented as mean ± SD. <sup>ns</sup> <span class="html-italic">p</span> &gt; 0.05, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001compared with the control group/ HC-GO. All experiments were independently repeated at least three times.</p>
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15 pages, 2445 KiB  
Article
Boswellia serrate Gum Resin Mitigates Renal Toxicity: Role of TNF-α, Interleukins, TGF-β, and Lipid Peroxidation
by Heba M. Eltahir, Abdel-Gawad S. Shalkami, Ahmed M. Shehata, Mohannad Almikhlafi, Ahmed J. Aldhafiri, Ali Alalawi, Muayad Albadrani, Ahmad Bakur Mahmoud and Mekky M. Abouzied
Life 2024, 14(12), 1669; https://doi.org/10.3390/life14121669 - 17 Dec 2024
Viewed by 255
Abstract
Background and aim: Being a central organ in homeostasis and maintaining the health of the biological system, kidneys are exposed to variable toxicants. Long-term exposure to nephrotoxic molecules causes chronic renal damage that causes fibrosis and loss of function. Such damage can be [...] Read more.
Background and aim: Being a central organ in homeostasis and maintaining the health of the biological system, kidneys are exposed to variable toxicants. Long-term exposure to nephrotoxic molecules causes chronic renal damage that causes fibrosis and loss of function. Such damage can be initiated by oxidative stress which provokes inflammation. We aim at investigating the potential therapeutic effects of Boswellia serrata (BS) gum resin extract in managing CCl4-induced renal toxicity. Methods: Male Wistar albino rats were assigned to groups: healthy control; CCl4-treated (CCl4, twice/week, for 6 weeks); CCl4 + BS-treated: CCl4 for 6 weeks followed by BS (150 mg/kg/day) for 2 weeks; and CCl4 + Silymarin-treated: CCl4 for 6 weeks followed by Silymarin (100 mg/kg/day) for 2 weeks. Blood and kidney tissue were utilized to assess oxidative stress status, inflammatory cytokines, and histopathological changes. Results: BS treatment ameliorated signs of renal damage and fibrosis as it improved renal antioxidant status and renal function markers and significantly reduced the levels of inflammatory cytokines TNF-α, IL-1β, IL-6, and IL-8 along with the fibrogenic marker TGF-β. Kidney tissues showed improved histological features after BS treatment. Conclusions: BS gum resin extract has significant therapeutic potential against CCl4-induced renal damage and fibrosis. These effects could be mediated via its previously reported antioxidant, free radical scavenging, and anti-inflammatory effects. Full article
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<p><b>Effect of the different treatments on TNF-α level in CCl<sub>4</sub>-induced renal toxicity.</b> CCl<sub>4</sub> administration resulted in a significant increase in TNF-α levels compared to healthy control animals. Treatment of animals with BS or SIL after the establishment of renal damage significantly ameliorated TNF-α levels compared to animals receiving CCl<sub>4</sub> without further treatment. Data are represented as mean ± SEM. *: values are significantly different from healthy control; #: values are significantly different from CCl<sub>4</sub>-treated animals. <span class="html-italic">p</span> &lt; 0.05.</p>
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<p><b>Effect of the different treatments on IL-1β, IL-6, and IL-8 levels in CCl<sub>4</sub>-induced renal toxicity.</b> CCl<sub>4</sub> administration resulted in a significant increase in the levels of all three interleukins IL-1β (<b>A</b>), IL-6 (<b>B</b>), and IL-8 (<b>C</b>) compared to healthy control animals. Treatment of animals with BS or SIL after establishment of renal damage significantly ameliorated the levels of these interleukins compared to animals receiving CCl<sub>4</sub> without further treatment. Data are represented as mean ± SEM. *: values are significantly different from healthy control; #: values are significantly different from CCl<sub>4</sub>-treated animals. <span class="html-italic">p</span> &lt; 0.05.</p>
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<p><b>Effect of the different treatments on TGF-β levels in CCl<sub>4</sub>-induced renal toxicity.</b> CCl<sub>4</sub> administration resulted in a significant increase in TGF-β levels compared to healthy control animals. Treatment of animals with BS or SIL after the establishment of renal damage significantly ameliorated TGF-β levels compared to animals receiving CCl<sub>4</sub> without further treatment. Data are represented as mean ± SEM. *: values are significantly different from healthy control; #: values are significantly different from CCl<sub>4</sub>-treated animals. <span class="html-italic">p</span> &lt; 0.05.</p>
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<p><b>Effect of the different treatments on renal histological architecture.</b> (<b>A</b>) Photomicrograph of kidney section from control animals showing normal renal architecture including normal renal corpuscles (R) and renal tubules (T) as well as intact, healthy blood vessels (V). (<b>B</b>) sections from renal tissue of CCl<sub>4</sub>-treated animals showing atrophy of the renal corpuscle (R) and severe degenerative changes in the renal tubules epithelium (T), as well as cystic dilatation and hyaline cast appearance (arrow). Infiltrating lymphocytes are visible in the interstitial tissue (arrowhead), and renal blood vessels (V) appear highly congested. (<b>C</b>) Renal tissue sections from CCl<sub>4</sub>-treated rats administered BS showing almost normal renal corpuscles (R) and renal tubules (T). Renal blood vessels (V) show signs of mild congestion. (<b>D</b>) Renal tissue sections from CCl<sub>4</sub>-treated rats administered Silymarin showing normal renal corpuscles (R) and renal tubules (T1); however, some tubules (T2) showed degenerative changes in their epithelia. Also, mildly congested renal blood vessels (V) could be observed. Hematoxylin and eosin staining; bars = 100 µm; 200×.</p>
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<p><b>Effect of the different treatments on ECM and collagen deposition.</b> (<b>A</b>) Photomicrograph of kidney section from control animals stained with Masson’s Trichrome stain showing no accumulation of collagen fibers. (<b>B</b>) Kidney sections from CCl<sub>4</sub>-treated group showing a strong positive staining for densely accumulated collagen fibers (arrows). (<b>C</b>,<b>D</b>) Kidney sections from BS- and SIL-treated animals, respectively, showing almost no accumulation of collagen fibers (Masson’s Trichrome stain, bars = 50 µm, and 400×).</p>
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<p><b>Effect of the different treatments on basement membrane integrity.</b> (<b>A</b>) A kidney section from the control group showing normal renal corpuscles (R) enclosed by intact basal lamina (arrow) and renal tubules (T) lined with tubular epithelium with normal apical brush borders (arrow head). (<b>B</b>) A kidney section from the CCl<sub>4</sub>-treated group showed partially lost brush borders (arrowhead), dilated renal tubules (T), and deformed basal lamina surrounding the renal corpuscle (arrow). (<b>C</b>) A kidney section from CCl<sub>4</sub>-treated rats administered BS showing intact basal lamina enclosing the renal corpuscle (R) (arrow); brush borders are partially lost in few renal tubules (T2), while the majority of tubules (T1) appeared normal with an apical brush border (arrow head). (<b>D</b>) A kidney section from CCl<sub>4</sub>-treated rats administered Silymarin showing a renal corpuscle (R) enclosed by a thick basal lamina (arrow) as well as a lost brush border and cystic dilatation in some renal tubules (T2), while the other tubules (T1) appeared normal with an apical brush border (arrow head). PAS staining; bars = 50 µm.</p>
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15 pages, 2430 KiB  
Article
Impact of Brake Wear Particles on Eukaryotic Cell Viability and Associated Oxidative Stress Responses
by Lina Trečiokaitė, Yurii Tsybrii, Oleksii Nosko and Lina Ragelienė
Lubricants 2024, 12(12), 449; https://doi.org/10.3390/lubricants12120449 - 16 Dec 2024
Viewed by 282
Abstract
In this study, the cytotoxic effects of brake wear particles (≥250 nm ceramic/ceramic wear particles (CCWPs) and ≤100 nm ceramic/steel wear particles (CSWPs)) and 100 nm iron (III) oxide ultrafine particles (IOUFPs) on human lung carcinoma (A549) and Chinese hamster ovary (CHO) cells [...] Read more.
In this study, the cytotoxic effects of brake wear particles (≥250 nm ceramic/ceramic wear particles (CCWPs) and ≤100 nm ceramic/steel wear particles (CSWPs)) and 100 nm iron (III) oxide ultrafine particles (IOUFPs) on human lung carcinoma (A549) and Chinese hamster ovary (CHO) cells were investigated. Cell viability was determined using the MTT and Calcein AM methods. Oxidative stress was assessed by measuring reactive oxygen species (ROS), intracellular reduced glutathione (GSH), and malondialdehyde (MDA) concentrations under exposure to the above particles in the concentration range of 10–80 µg/mL. The initial assessments of CCWPs and CSWPs on the cell viability were performed after a 4-h exposure but later extended to 24 h to investigate the time-dependent of the cell viability and oxidative stress. MTT and Calcein AM assays indicated that the A549 cells are less susceptible to CCWPs and CSWPs than the CHO cells when exposed for both 4 h and 24 h. This study highlights that oxidative stress induced by CCWPs, CSWPs, and IOUFPs is cell-specific. While CCWPs did not affect glutathione (GSH) levels in the CHO cells, it significantly reduced GSH levels in A549 cells, with the exception of 80 µg/mL. Both CCWPs and CSWPs increased the lipid peroxidation in both cell types; however, the A549 cells demonstrated lower sensitivity to these treatments. Full article
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Figure 1
<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspensions for 4 h on the viability of CHO and A549 cells determined by the MTT method. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspensions for 4 h on the viability of CHO and A549 cells determined by the Calcein AM method. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspensions for 24 h on the viability of CHO and A549 cells determined by the MTT method. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspensions for 24 h on the viability of CHO and A549 cells determined by the Calcein AM method. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspension for 24 h on the ROS formation in the CHO and A549 cells. CHO had 30,000 fluorescence intensity (<b>A</b>,<b>B</b>), and A459 had 28,000 fluorescence intensity (<b>A</b>,<b>B</b>), with 100 µM H<sub>2</sub>O<sub>2</sub>. Blank means cells without H<sub>2</sub>O<sub>2</sub>, CCWPs, CSWPs, and IOUFPs. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspension for 24 h on the GSH of CHO and A549 cells. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Effect of the CCWP, CSWP (<b>A</b>), and IOUFP (<b>B</b>) suspensions for 24 h on the lipid peroxidation of CHO and A549 cells. Data are mean ± SD of at least 3 replicates, <span class="html-italic">n</span> = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001 compared to the control group (viability of cells without exposure of particles 100%): (a) significant difference as compared with CCWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; (b) significant difference as compared with CSWP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05; and (c) significant difference as compared with IOUFP between cell CHO and A549 lines at <span class="html-italic">p</span> &lt; 0.05.</p>
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22 pages, 5873 KiB  
Article
Quality by Design Approach for the Formulation and Evaluation of Stem Cells Derived Rosmarinic Acid-Loaded Nanofibers as an Anti-Wrinkle Patch: In Vitro and In Vivo Characterizations
by Rehab Abdelmonem, Ahmed Bakr, Ingy Badawy, Ahmed Ibrahim Abd El Maksoud and Reem T. Attia
Pharmaceutics 2024, 16(12), 1598; https://doi.org/10.3390/pharmaceutics16121598 - 16 Dec 2024
Viewed by 313
Abstract
Background/Objectives: Skin wrinkles result from a myriad of multifaceted processes involving intrinsic and extrinsic aging. To combat this effect, plant stem cells offer a renewable and eco-friendly source for various industries, including cosmeceuticals. Salvia miltiorrhiza (SM), which contains the bioactive compound Rosmarinic acid [...] Read more.
Background/Objectives: Skin wrinkles result from a myriad of multifaceted processes involving intrinsic and extrinsic aging. To combat this effect, plant stem cells offer a renewable and eco-friendly source for various industries, including cosmeceuticals. Salvia miltiorrhiza (SM), which contains the bioactive compound Rosmarinic acid (RA) and has been proposed for its anti-wrinkle effect. Methods: In the present study, calli from SM were cultured and Quality by Design (QbD) was implemented to investigate the effect of different types and concentrations of elicitors; jasmonic acid (JA) and salicylic acid (SA). Both raised RA levels yet, jasmonic acid (50 µM) has resulted in the highest yield for RA, at 16 mg/g. A nanofiber patch was prepared and characterized in-vitro by the release percentage, drug content, swelling degree, scanning electron microscope, and surface roughness. Then, the anti-wrinkle effect of the patch was tested in a UV wrinkle-induced mouse model. Results: Interestingly, after treatment, there were visibly fewer wrinkles, and the skin was softer than in the untreated control group. This study suggests that the treatment exerted its effect through the Nrf2/Keap1 pathway, which plays a crucial role in cellular antioxidant protective processes. By activating this pathway through boosting Nrf2 and diminishing Keap1 cellular content, the nanofiber patch enhances the production of antioxidant enzymes, such as superoxide dismutase and glutathione peroxidase, enhancesglutathione, and reduces the skin lipid peroxidation, collectively indicating enhanced skin quality. Conclusions: In conclusion, this study highlights the importance of this formula as an anti-wrinkle treatment, and future clinical studies are recommended to further unveil the potential of this formula. Full article
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<p>A chart showing the effect of different concentrations of Jasmonic acid and Salicylic acid on Rosmarinic acid (RA) yield in the <span class="html-italic">Salvia miltiorrhiza</span> stem cell extract compared with the (0) control: RA yield without elicitors, which is indicated with the green column.</p>
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<p>The effect of Jasmonic acid and Salicylic acid elicitor concentrations on Rosmarinic acid level, solid lines represent the predicted responses from the statistical model and dashed lines Indicate observed experimental responses.</p>
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<p>Desirability ramps for the ideal elicitor concentration where red dots represent jasmonic acid concentration and blue dots represent rosmarinic acid concentration.</p>
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<p>In vitro release profile of Rosmarinic acid from nanofiber patch in acetate buffer pH 5.5. Data are presented as mean+/− SD at <span class="html-italic">p</span> &lt; 0.05 <span class="html-italic">n</span> = 3.</p>
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<p>FTIR spectrum of Polyurethane/Rosmarinic acid electrospun nanofiber.</p>
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<p>In vitro release profile of Rosmarinic acid electrospun nanofiber in acetate buffer pH 5.5 showing the patch’s stability after 3 months of preparation. Data are presented as mean+/− SD at <span class="html-italic">p</span> &lt; 0.05 <span class="html-italic">n</span> = 3.</p>
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<p>SEM images of electrospun RA/PU nanofiber patch, polymer solution 10%wt with flow rate 0.8 mL/h and 25 kV, (<b>A</b>) 30 µM, (<b>B</b>) 10 µM, (<b>C</b>) Free Polyurethane. Green arrows sign to the nanospheres of extracted rosmarinic acid, yellow arrows sign to the uniformly loaded nanofibers.</p>
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<p>Surface roughness images for Rosmarinic acid/Polyurethane nanofiber patch.</p>
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<p>Photograph of a sample of the mice’s back after treatment. (<b>A</b>) Normal control (NC) group, (<b>B</b>) positive control (PC) group, (<b>C</b>) free patch group (Free-P), (<b>D</b>) liquid RA group, (<b>E</b>) medicated nanofiber patch-treated group (Mediated P).</p>
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<p>Skin histopathological images. (<b>A</b>) Normal control (NC) group, (<b>B</b>) positive control (PC) group, (<b>C</b>) free patch group (Free-P), (<b>D</b>) liquid RA group, (<b>E</b>) medicated nanofiber patch-treated group (Medicated P). Arrows represent the variation in epidermal thickening and the presence of necrotic tissues.</p>
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<p>Effect of different treatments on skin Nrf2 and Keap1 content. Data are presented as mean ± SD (<span class="html-italic">n</span> = 6). Statistical analysis was carried out using one-way ANOVA followed by Tukey’s Multiple Comparison test, and the used <span class="html-italic">p</span> value is shown for each comparison in the graph. Each shape over the bar-chart represents a sample that was tested.</p>
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<p>Effect of different treatments on skin GSH and GPx, SOD, MDA. Data are presented as mean ± SD (<span class="html-italic">n</span> = 6). Statistical analysis was carried out using one-way ANOVA followed by Tukey’s Multiple Comparison test, and the used <span class="html-italic">p</span> value is shown for each comparison in the graph. Each shape over the bar-chart represents a sample that was tested.</p>
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20 pages, 2978 KiB  
Article
Response of Arabidopsis thaliana to Flooding with Physical Flow
by Momoko Kaji, Kazuma Katano, Taufika Islam Anee, Hiroshi Nitta, Ryotaro Yamaji, Rio Shimizu, Shunsuke Shigaki, Hiroyuki Suzuki and Nobuhiro Suzuki
Plants 2024, 13(24), 3508; https://doi.org/10.3390/plants13243508 - 16 Dec 2024
Viewed by 276
Abstract
Flooding causes severe yield losses worldwide, making it urgent to enhance crop tolerance to this stress. Since natural flooding often involves physical flow, we hypothesized that the effects of submergence on plants could change when combined with physical flow. In this study, we [...] Read more.
Flooding causes severe yield losses worldwide, making it urgent to enhance crop tolerance to this stress. Since natural flooding often involves physical flow, we hypothesized that the effects of submergence on plants could change when combined with physical flow. In this study, we analyzed the growth and transcriptome of Arabidopsis thaliana exposed to submergence or flooding with physical flow. Plants exposed to flooding with physical flow had smaller rosette diameters, especially at faster flow rates. Transcriptome analysis revealed that “defense response” transcripts were highly up-regulated in response to flooding with physical flow. In addition, up-regulation of transcripts encoding ROS-producing enzymes, SA synthesis, JA synthesis, and ethylene signaling was more pronounced under flooding with physical flow when compared to submergence. Although H2O2 accumulation changed in response to submergence or flooding with physical flow, it did not lead to lipid peroxidation, suggesting a role for ROS as signaling molecules under these conditions. Multiple regression analysis indicated possible links between rosette diameter under flooding with physical flow and the expression of Rbohs and SA synthesis transcripts. These findings suggest that pathogen defense responses, regulated by SA and ROS signaling, play crucial roles in plant responses to flooding with physical flow. Full article
(This article belongs to the Special Issue Deciphering Plant Molecular Data Using Computational Methods)
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<p>Schematic design of the channel to treat plants with flooding. (<b>A</b>) Top view, (<b>B</b>) side view.</p>
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<p>Growth characteristics of plants subjected to submergence or flooding with physical flow. (<b>A</b>) Number of leaves, (<b>B</b>) plant diameter and (<b>C</b>) inflorescent length. Error bars indicate standard deviation (<span class="html-italic">n</span> = 9–10). * and **: Student’s <span class="html-italic">t</span>-test significantly different at <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01, respectively (compared to control).</p>
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<p>Characteristics of transcripts up-regulated in response to flooding with physical flow. (<b>A</b>) Venn diagram showing the overlap between transcripts up-regulated in response to submergence or flooding with physical flow. (<b>B</b>,<b>C</b>) Gene Ontology (GO) terms of “biological processes” represented in transcripts specifically up-regulated in response to submergence (<b>B</b>) or flooding with physical flow (<b>C</b>).</p>
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<p>Involvement of pathogen defense pathways in response of plants to flooding with physical flow. (<b>A</b>) Fold change of transcripts belong to Gene Ontologies (GOs) that are highly represented in the transcripts specifically up-regulated in response to flooding with physical flow. (<b>B</b>) Proportion of hormone response transcripts among the transcripts specifically up-regulated in response to submergence or flooding with physical flow. ABA: abscisic acid, ACC: 1-aminocyclopropane-1-carboxylate, BL: brassinosteroids, CK: Cytokinin, GA: gibberellic acid, IAA: indole-3-acetic acid, MJ: methyl jasmonate, SA: salicylic acid.</p>
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<p>FPKM of transcripts involved in Salicylic acid (SA) synthesis (<b>A</b>), Jasmonic acid (JA) synthesis (<b>B</b>) or ROS production (<b>C</b>). Values relative to control are indicated. Error bars indicate standard deviation (<span class="html-italic">n</span> = 3). * and **: Student’s <span class="html-italic">t</span>-test significantly different at <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01, respectively (compared to control).</p>
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<p>FPKM of transcripts encoding ERFs. Values relative to control are indicated. Error bars indicate standard deviation (<span class="html-italic">n</span> = 3). Transcripts were categorized into three groups; Sub: significantly up-regulated specifically in response to submergence, Flood: significantly up-regulated specifically in response to flooding with physical flow, Sub and Flood: significantly up-regulated in response to both stresses.</p>
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<p>Multiple regression analysis of transcripts involved in ROS production ((<b>A</b>) <span class="html-italic">Rboh</span>s), SA (salicylic acid) synthesis (<b>B</b>) and AU (auxin) signaling (<b>C</b>). Circles indicate the transcript that showed the highest contribution to the determination of rosette diameter under flooding with physical flow. In this figure, the <span class="html-italic">X</span>- and <span class="html-italic">Y</span>-axis values indicate the ranking of transcripts that contribute to the effects on rosette diameter, with lower numbers indicating a higher contribution.</p>
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<p>Accumulation of H<sub>2</sub>O<sub>2</sub> (<b>A</b>) and MDA (<b>B</b>) in plants exposed to submergence (Sub) or flooding with different flow rates. Error bars indicate standard deviation (<span class="html-italic">n</span> = 3). * and **: Student’s <span class="html-italic">t</span>-test significantly different at <span class="html-italic">p</span> &lt; 0.05 and <span class="html-italic">p</span> &lt; 0.01, respectively (<span class="html-italic">n</span> = 3, compared to control).</p>
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17 pages, 5770 KiB  
Article
Genome-Wide Identification of the SlSET Gene Family and the Function of SlSET6 Under Salt Stress
by Xueying Yang, Yan Gao, Chengyu Zhu, Xin Li, Yuliang Gao and Kuihua Li
Int. J. Mol. Sci. 2024, 25(24), 13461; https://doi.org/10.3390/ijms252413461 - 16 Dec 2024
Viewed by 273
Abstract
A comprehensive genome-wide identification of SET-domain-containing genes in Solanum lycopersicum (tomato) has revealed 46 members. Phylogenetic analysis showed that these SET genes, along with those from Arabidopsis thaliana and Oryza sativa, are divided into five subfamilies, with Subfamilies II and V being [...] Read more.
A comprehensive genome-wide identification of SET-domain-containing genes in Solanum lycopersicum (tomato) has revealed 46 members. Phylogenetic analysis showed that these SET genes, along with those from Arabidopsis thaliana and Oryza sativa, are divided into five subfamilies, with Subfamilies II and V being the largest. Motif and domain analyses identified 15 conserved motifs and revealed the presence of pre-SET and post-SET domains in several genes, suggesting functional diversification. Gene structure analysis further demonstrated variation in exon–intron organization, likely contributing to differential gene regulation. Promoter analysis identified cis-acting elements related to light responsiveness, plant growth, hormones, and stress, implicating SET genes in various biological processes. RNA-seq and qRT-PCR data revealed distinct expression patterns of SlSET genes under salt stress, with several genes showing significant upregulation, indicating their potential role in stress tolerance. In particular, SlSET6 silencing using VIGS reduced tomato’s tolerance to salt stress, leading to higher lipid peroxidation, reduced antioxidant enzyme activity, and decreased proline content, further confirming its critical role in salt stress response. These findings provide valuable insights into the functional diversity, evolutionary history, and stress-related roles of SET domain genes in tomato, with potential applications for crop improvement strategies. Full article
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<p>Phylogenetic analysis of SET proteins from Arabidopsis, rice, and tomato. Five different subfamilies are represented by red, purple, blue, orange, and pink, respectively. Circles, triangles, and stars represent Arabidopsis, rice, and tomato.</p>
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<p>Phylogenetic, motif, conserved domain, and structural analyses. (<b>a</b>) Phylogenetic tree of SlSET proteins. (<b>b</b>) Conserved motifs of SlSET proteins: a total of 15 motifs were identified; different colored squares indicate different motifs. (<b>c</b>) Conserved domains of SlSET proteins: a total of 4 types of SET domains were identified. (<b>d</b>) Gene structure of <span class="html-italic">SET</span> genes.</p>
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<p><span class="html-italic">Cis</span>-element analysis of SET genes’ promoters. <span class="html-italic">Cis</span>-acting elements are divided into 4 categories according to their functions, including light responsiveness, plant growth and development, plant hormone related and stress related. The number represents the number of <span class="html-italic">cis</span>-acting elements in the promoter region of the corresponding gene.</p>
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<p>Location and collinearity analysis of <span class="html-italic">SET</span> genes. Red lines and red names indicate <span class="html-italic">SlSET</span> genes that have a collinearity relationship. Gray lines indicate all the collinearity genes in the tomato genome.</p>
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<p>Synteny analysis of <span class="html-italic">SET</span> genes in <span class="html-italic">S. lycopersicum</span> (orange), <span class="html-italic">A. thaliana</span> (green), <span class="html-italic">O. sativa</span> (yellow), and <span class="html-italic">S. tuberosum</span> (brown). Gray lines indicate genes have a synteny relationship in different genomes, and blue lines indicate <span class="html-italic">SET</span> genes.</p>
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<p>Transcriptomic heatmap of <span class="html-italic">SlSET</span> genes under salt stress. Red color indicates a high expression level under salt stress; white color indicates a low expression level under salt stress. A 1, 2 and 3 at the end of samples denotes the biological replicates. The heatmap was visualized using TBtools software (v2.142).</p>
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<p>qRT-PCR analysis of 9 <span class="html-italic">SlSET</span> genes under salt stress. Error bars on the graph represent the standard error of the mean (SEM; n = 3 biological replicates). Different letters indicate statistically significance differences between groups, as determined using Fisher’s LSD test at a 5% level of significance. Experimental data were subjected to a one-way ANOVA.</p>
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<p>Functional analysis of the <span class="html-italic">SlSET6</span> gene under salt stress. (<b>a</b>) Phenotypic observations of pTRV2-00 and pTRV2-<span class="html-italic">SlSET6</span> plants. (<b>b</b>) Relative expression level of the <span class="html-italic">SlSET6</span> gene in pTRV2-00 and pTRV2-<span class="html-italic">SlSET6</span> plants. (<b>c</b>) MDA content. (<b>d</b>) CAT activity. (<b>e</b>) Pro content. (<b>f</b>) POD activity. (<b>g</b>) SOD activity. Error bars on the graph represent the standard error of the mean (SEM; n = 3 biological replicates). Different letters indicate statistically significance differences between groups, as determined using Fisher’s LSD test at a 5% level of significance. Experimental data were subjected to a one-way ANOVA.</p>
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14 pages, 14355 KiB  
Article
JP4-039 Mitigates Cisplatin-Induced Acute Kidney Injury by Inhibiting Oxidative Stress and Blocking Apoptosis and Ferroptosis in Mice
by Merlin Airik, Kacian Clayton, Peter Wipf and Rannar Airik
Antioxidants 2024, 13(12), 1534; https://doi.org/10.3390/antiox13121534 - 15 Dec 2024
Viewed by 388
Abstract
Cisplatin is a commonly used chemotherapeutic agent in the treatment of a wide array of cancers. Due to its active transport into the kidney proximal tubule cells, cisplatin treatment can cause a buildup of this nephrotoxic compound in the kidney, resulting in acute [...] Read more.
Cisplatin is a commonly used chemotherapeutic agent in the treatment of a wide array of cancers. Due to its active transport into the kidney proximal tubule cells, cisplatin treatment can cause a buildup of this nephrotoxic compound in the kidney, resulting in acute kidney injury (AKI). About 30% of patients receiving cisplatin chemotherapy develop cisplatin-induced AKI. JP4-039 is a mitochondria-targeted reactive oxygen species (ROS) and electron scavenger. Recent studies have shown that JP4-039 mitigates a variety of genotoxic insults in preclinical studies in rodents by suppressing oxidative stress-mediated tissue damage and blocking apoptosis and ferroptosis. However, the benefits of JP4-039 treatment have not been tested in the setting of AKI. In this study, we investigated the potential renoprotective effect of JP4-039 on cisplatin-induced AKI. To address this goal, we treated mice with JP4-039 before or after cisplatin administration and analyzed them for functional and molecular changes in the kidney. JP4-039 co-administration attenuated cisplatin-induced renal dysfunction and histopathological changes. Upregulation of tubular injury markers was also suppressed by JP4-039. Mechanistically, JP4-039 suppressed lipid peroxidation, prevented tissue oxidative stress, and preserved the glutathione levels in cisplatin-injected mice. An increase in cisplatin-induced apoptosis and ferroptosis was also alleviated by the compound. Moreover, JP4-039 inhibited cytokine overproduction in cisplatin-injected mice. Together, our findings demonstrate that JP4-039 is a promising therapeutic agent against cisplatin-induced kidney injury. Full article
(This article belongs to the Special Issue Oxidative Stress in Renal Health)
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<p>Cisplatin-induced kidney injury and loss of function is mitigated by JP4-039. (<b>A</b>) Overview of the cisplatin AKI and JP4-039 administration protocols. Cisplatin was administered to three cohorts of 129Sv-Elite mice; cisplatin cohort (<b>C</b>) received a single dose of cisplatin (10 mg/kg), cohort 2 (C + JP4-039) received a single dose of cisplatin (10 mg/kg), followed by JP4-039 (20 mg/kg) 24 h later, and cohort 3 (JP4-039 + C) was injected with JP4-039 (20 mg/kg) followed by cisplatin (10 mg/kg) 1 h later. Control mice (ctrl) were injected with normal saline and DMSO (vehicles for cisplatin and JP4-039, respectively). Kidneys and blood were collected for analysis 3 days after cisplatin injection. (<b>B</b>) Histological analysis of kidney sections via periodic acid–Schiff (PAS) staining reveals that JP4-039 treatment preserves the kidneys from cisplatin injury. Green asterisks, protein casts; arrowheads, loss of brush-border. Scale bars: 100 μm. (<b>C</b>) Blood urea nitrogen (BUN) measurements demonstrate AKI in cisplatin-only but not in control and JP4-039-treated mice. Ordinary one-way ANOVA with Tukey’s multiple comparison ** <span class="html-italic">p</span> &lt; 0.01, **** <span class="html-italic">p</span> &lt; 0.0001, <span class="html-italic">n</span> = 5 each. (<b>D</b>) Tubular injury scores based on PAS-stained kidney sections. Ordinary one-way ANOVA with Tukey’s multiple comparison ** <span class="html-italic">p</span> &lt; 0.01, <span class="html-italic">n</span> = 5 each. (<b>C</b>,<b>D</b>) Data are presented as the mean ± SEM. A two-way ANOVA with Tukey’s post hoc analysis.</p>
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<p>JP4-039 administration mitigated tubular injury in cisplatin AKI. (<b>A</b>) Representative images of Kim1 immunofluorescence staining in control (untreated) and cisplatin ± JP4-039-treated kidneys on day 3 after AKI induction, Kim1 (red), LTL (green), DAPI (blue). Scale bar: 200 μm. (<b>B</b>) Quantification of KIM1-positive area in LTL-positive proximal tubules shows that prophylactic and therapeutic JP4-039 administrations suppresses KIM1 expression in cisplatin-injected kidneys. One way-ANOVA, ** <span class="html-italic">p</span> &lt; 0.01, <span class="html-italic">n</span> = 5 each. (<b>C</b>) <span class="html-italic">Havcr1</span> expression was increased in cisplatin AKI kidneys but remained at baseline in mice co-administered with JP4-039. One way-ANOVA, *** <span class="html-italic">p</span> &lt; 0.001, <span class="html-italic">n</span> = 5 each. (<b>D</b>) <span class="html-italic">Ngal</span> expression was increased in cisplatin AKI kidneys but remained at baseline in the kidneys of mice co-administered JP4-039. One way-ANOVA, ** <span class="html-italic">p</span> &lt; 0.01, <span class="html-italic">n</span> = 5 each. (<b>B</b>–<b>D</b>) Data are presented as the mean ± SEM. One way-ANOVA with Tukey’s post hoc analysis.</p>
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<p>JP4-039 blocked the development of tubular interstitial fibrosis in cisplatin AKI. (<b>A</b>) Representative images of Collagen1 immunofluorescence staining in control (untreated) and cisplatin ± JP4-039-treated kidneys on day 3 after AKI induction; collagen 1a1 (red), LTL (green), and DAPI (blue). Scale bar: 200 μm. (<b>B</b>) Quantification of collagen 1-positive area in the cortical region shows that prophylactic and therapeutic JP4-039 administrations suppress Col1a expression in cisplatin-injected kidneys. One way-ANOVA, **** <span class="html-italic">p</span> &lt; 0.0001, <span class="html-italic">n</span> = 5 each. (<b>C</b>) Representative images of fibronectin 1 immunofluorescence staining in control (untreated) and cisplatin ± JP4-039-treated kidneys on day 3 after AKI induction; fibronectin 1 (red), LTL (green), DAPI (blue). Scale bar: 200 μm. (<b>D</b>) Quantification of fibronectin 1-positive area in the cortical region shows that prophylactic and therapeutic JP4-039 treatments suppress Fn1 expression in cisplatin-injected kidneys. (<b>B</b>,<b>D</b>) Data are presented as the mean ± SEM. One way-ANOVA, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001, <span class="html-italic">n</span> = 5 each.</p>
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<p>JP4-039 treatment reduced intrarenal infiltration of inflammatory cells triggered by tubular cell death. (<b>A</b>–<b>D</b>) Evaluation of the expression of several inflammatory genes—<span class="html-italic">Il6</span> (<b>A</b>), <span class="html-italic">Cxcl10</span> (<b>B</b>), <span class="html-italic">Tnf</span> (<b>C</b>), and <span class="html-italic">Ccl2</span> (<b>D</b>) by qPCR reveals a robust inflammatory signaling in cisplatin AKI, which is suppressed in mice co-treated with JP4-039. (<b>A</b>–<b>D</b>) Data are presented as the mean ± SEM. One way-ANOVA with Tukey’s post hoc analysis, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, <span class="html-italic">n</span> = 5 per group.</p>
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<p>JP4-039 blocked the generation of reactive oxygen species (ROS) and activation of the Nrf2 antioxidant pathway in cisplatin-treated kidneys. (<b>A</b>) CM-H2DCFDA staining in untreated and cisplatin ± JP4-039-treated kidneys. Scale bar: 200 μm. (<b>B</b>) Quantification of the CM-H2DCFDA. **** <span class="html-italic">p</span> &lt; 0.0001, n = 5. (<b>C</b>–<b>F</b>) Expression of <span class="html-italic">Nrf2</span> (<b>C</b>) and its target genes, <span class="html-italic">Nqo1</span> (<b>D</b>) and <span class="html-italic">Hmox</span> (<b>E</b>), was increased in cisplatin AKI kidneys but remained at baseline in mice co-administered with JP4-039, while <span class="html-italic">Gpx6</span> (<b>F</b>) expression was reduced in AKI kidneys but preserved at baseline in JP4-039 administered kidneys. One way-ANOVA, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001, <span class="html-italic">n</span> = 5 each. (<b>B</b>–<b>F</b>) Data are presented as the mean ± SEM. One way-ANOVA with Tukey’s post hoc analysis.</p>
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<p>JP4-039 reduced apoptosis and ferroptosis in tubular epithelial cells in cisplatin-injured kidneys. (<b>A</b>) Representative images of cleaved Caspase-3 immunohistochemistry staining in untreated and cisplatin ± JP4-039-treated kidneys. Scale bar: 100 μm. (<b>B</b>) Quantification of cleaved Caspase-3 positive cells in the kidney cortex. ** <span class="html-italic">p</span> &lt; 0.01, <span class="html-italic">n</span> = 5 per group. (<b>C</b>) 4-Hydroxynonenal (4-HNE) staining in control and AKI ± JP4-039 kidneys showed that JP4-039 mitigated the cisplatin-induced accumulation of lipid peroxidation in kidneys 3 days after cisplatin administration. Scale bar: 100 μm. (<b>D</b>) Quantification of 4HNE positive area in the kidney cortical area. **** <span class="html-italic">p</span> &lt;0.0001, <span class="html-italic">n</span> = 5 per group. (<b>E</b>) Quantification of free GSH concentration in whole kidney lysates. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, <span class="html-italic">n</span> = 3 per group. (<b>F</b>,<b>G</b>) qPCR analysis of <span class="html-italic">Acsl4</span> and <span class="html-italic">Gpx4</span> expression in kidneys normalized to <span class="html-italic">Gapdh,</span> * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, <span class="html-italic">n</span> = 5 per group. (<b>H</b>,<b>I</b>) Western blot and quantification of kidney Acsl4 protein. Tubulin is a loading control. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt;0.0001, <span class="html-italic">n</span> = 5 per group. (<b>B</b>,<b>D</b>–<b>F</b>,<b>G</b>,<b>I</b>) Data are presented as the mean ± SEM. One way-ANOVA with Tukey’s post hoc analysis.</p>
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15 pages, 5153 KiB  
Article
Impact of Arsenic Stress on the Antioxidant System and Photosystem of Arthrospira platensis
by Jiawei Liu, Jie Du, Di Wu, Xiang Ji and Xiujuan Zhao
Biology 2024, 13(12), 1049; https://doi.org/10.3390/biology13121049 - 15 Dec 2024
Viewed by 363
Abstract
Arthrospira platensis exhibits high tolerance to arsenic; however, the mechanisms underlying its response to the arsenic stress have not been fully elucidated. This study investigated the growth and resistance mechanisms of A. platensis under As3+ stress by measuring physiological and biochemical indices, [...] Read more.
Arthrospira platensis exhibits high tolerance to arsenic; however, the mechanisms underlying its response to the arsenic stress have not been fully elucidated. This study investigated the growth and resistance mechanisms of A. platensis under As3+ stress by measuring physiological and biochemical indices, conducting transcriptome sequencing, and validating the results through qPCR. The findings show that arsenic stress affected the antioxidant system and photosynthetic pigment synthesis in A. platensis. The algae mitigated arsenic-induced oxidative stress by increasing cellular metabolic rates, enhancing cell wall stability, and reducing membrane lipid peroxidation. Transcriptome analysis revealed that pathways related to oxidative phosphorylation and chlorophyll degradation were upregulated under arsenic stress, while the expression of membrane transporters was significantly downregulated. Additionally, the algae alleviated arsenic stress by producing hydrogen and polyamine compounds. This study provides insights into the mechanisms of A. platensis response to arsenic stress and elucidates the molecular pathways involved in the stress response to As3+. Full article
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<p>Effect of As<sup>3+</sup> on the growth and biological activity concentration of <span class="html-italic">A. platensis</span>. (<b>a</b>) Growth of <span class="html-italic">A. platensis</span>. (<b>b</b>) PC content. (<b>c</b>) Carotenoids content. (<b>d</b>) Chlorophyll a content. Different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The photosynthetic system of <span class="html-italic">A. platensis</span>. (<b>a</b>) Results of OJIP fluorescence kinetics curves. (<b>b</b>) Chlorophyll fluorescence transients (Fv/Fm, ETo/RC, TRo/RC, ABS/CS, ABS/RC, Sm, Vj, Mo, Wk, and Plabs).</p>
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<p>Effects of As<sup>3+</sup> stress on the antioxidant system of <span class="html-italic">A. platensis</span>. (<b>a</b>) SOD activities. (<b>b</b>) CAT activities. (<b>c</b>) MDA content. Different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Transcriptome sequencing results. (<b>a</b>) Cluster analysis between samples. In the figure, the right and lower sides represent sample names, the left and upper sides are sample clustering, and the squares with different colors represent the correlation between the two samples (<b>b</b>) Hierarchically clustered heatmap of gene expression. Each column in the figure represents a sample, and each row represents a gene. The color in the heat map indicates the expression amount of the gene in the sample. See the number annotation under the color bar at the upper right for the specific expression amount change trend. (<b>c</b>) Volcano plot of differently expressed genes with <span class="html-italic">p</span> &lt; 0.05 and |log2FC| ≥ 0.875.</p>
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<p>Results of KEGG enrichment analysis. The abscissa is the name of KEGG metabolic pathway, and the ordinate is the number of genes annotated to the pathway. See the legend for functional classification and grouping information. (<b>a</b>) KEGG enrichment analysis for all differentially expressed genes. (<b>b</b>) KEGG enrichment analysis for upregulated differentially expressed genes. (<b>c</b>) KEGG enrichment analysis for downregulated differentially expressed genes.</p>
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<p>Results of GO functional enrichment analysis. The vertical axis represents GO term, and the horizontal axis represents rich factor [refers to the ratio of the number of genes annotated to the GO term in the gene set to the number of genes annotated to the GO term. The larger the rich factor, the greater the degree of enrichment]. The size of the point represents the number of genes/transcripts in this go term, and the color of the point corresponds to different FDR ranges. (<b>a</b>) GO enrichment analysis for all differentially expressed genes. (<b>b</b>) GO enrichment analysis for upregulated differentially expressed genes. (<b>c</b>) GO enrichment analysis for downregulated differentially expressed genes.</p>
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<p>Oxidative phosphorylation pathway under As<sup>3+</sup> stress. (<b>a</b>) Oxidative phosphorylation pathway. ( Red indicates upregulation, while blue indicates downregulation.) (<b>b</b>) qPCR results of the key genes of the oxidative phosphorylation. (** indicate significant differences (<span class="html-italic">p</span> &lt; 0.05))</p>
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<p>Phaeophytin a metabolism pathway under As<sup>3+</sup> stress. (<b>a</b>) Phaeophytin a metabolism pathway. (the arrow indicates upregulation) (<b>b</b>) qPCR results of the key genes of the photosynthetic system. (* indicate significant differences (<span class="html-italic">p</span> &lt; 0.05))</p>
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<p>qPCR results. (<b>a</b>) qPCR result of the gene. (<b>b</b>) qPCR result of <span class="html-italic">MgtC family protein</span> gene. (* indicate significant differences (<span class="html-italic">p</span> &lt; 0.05))</p>
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23 pages, 1803 KiB  
Review
Tumor Metabolic Reprogramming and Ferroptosis: The Impact of Glucose, Protein, and Lipid Metabolism
by Keyu Zhu, Yuang Cai, Lan Lan and Na Luo
Int. J. Mol. Sci. 2024, 25(24), 13413; https://doi.org/10.3390/ijms252413413 - 14 Dec 2024
Viewed by 264
Abstract
Ferroptosis, a novel form of cell death discovered in recent years, is typically accompanied by significant iron accumulation and lipid peroxidation during the process. This article systematically elucidates how tumor metabolic reprogramming affects the ferroptosis process in tumor cells. The paper outlines the [...] Read more.
Ferroptosis, a novel form of cell death discovered in recent years, is typically accompanied by significant iron accumulation and lipid peroxidation during the process. This article systematically elucidates how tumor metabolic reprogramming affects the ferroptosis process in tumor cells. The paper outlines the basic concepts and physiological significance of tumor metabolic reprogramming and ferroptosis, and delves into the specific regulatory mechanisms of glucose metabolism, protein metabolism, and lipid metabolism on ferroptosis. We also explore how complex metabolic changes in the tumor microenvironment further influence the response of tumor cells to ferroptosis. Glucose metabolism modulates ferroptosis sensitivity by influencing intracellular energetic status and redox balance; protein metabolism, involving amino acid metabolism and protein synthesis, plays a crucial role in the initiation and progression of ferroptosis; and the relationship between lipid metabolism and ferroptosis primarily manifests in the generation and elimination of lipid peroxides. This review aims to provide a new perspective on how tumor cells regulate ferroptosis through metabolic reprogramming, with the ultimate goal of offering a theoretical basis for developing novel therapeutic strategies targeting tumor metabolism and ferroptosis. Full article
(This article belongs to the Special Issue Cancer Cell Metabolism: New Advances and Potential Therapies)
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<p>Ferroptosis mechanism: Ferroptosis is directly triggered by lipid peroxides, with iron ions catalyzing the Fenton reaction to generate reactive oxygen species (ROS) that lead to the formation of lipid peroxides. Polyunsaturated fatty acids (PUFAs) and arachidonic acid (AA) in the cell membrane can undergo a series of catalytic reactions to form peroxidized lipids, ultimately resulting in ferroptosis. Glutathione peroxidase 4 (GPX4) plays a pivotal role in inhibiting ferroptosis by consuming glutathione (GSH) to scavenge ROS. The Xc-system facilitates GSH synthesis by transporting glutamate and cystine, thereby suppressing ferroptosis. Ferroptosis inducers (FINs) promote ferroptosis by inhibiting both the Xc-system and GPX4. Ubiquinone (CoQ10), under the catalysis of ferroptosis suppressor protein 1 (FSP1), can be converted into its reduced form, CoQ10H2, which scavenges ROS and thus inhibits ferroptosis. (Abbreviations: SLC7A11, solute carrier family 7 member 11; SLC3A2, solute carrier family 3 member 2; PPP, pentose phosphate pathway; ACSL4, acyl-CoA synthetase long-chain family member 4; LPCAT3, lysophosphatidylcholine acyltransferase 3; ALOX15, arachidonate 15-lipoxygenase.) Created in <a href="http://BioRender.com" target="_blank">BioRender.com</a> (accessed on 9 September 2024).</p>
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<p>Carbohydrate metabolism and ferroptosis: (<b>A</b>) In normal cells, glucose (Glu) primarily undergoes complete oxidative decomposition. Within the electron transport chain of the mitochondrial inner membrane, electrons from complex I and complex II can occasionally transfer to oxygen molecules, generating reactive oxygen species (ROS) that induce ferroptosis. Concurrently, coenzyme Q (CoQ) in the electron transport chain scavenges some ROS, thereby inhibiting ferroptosis. (<b>B</b>) In tumor cells, Glu metabolism is primarily glycolytic. Hypoxia-inducible factor 1 subunit alpha (HIF-1α) promotes the expression of glucose transporters (GLUT), glycolytic enzymes, and lactate metabolism enzymes, thereby enhancing glycolysis in tumor cells. Consequently, the formation of ROS through complete glucose oxidation in the electron transport chain is reduced, leading to a decrease in ferroptosis. Additionally, tumor cells shunt more glucose into the pentose phosphate pathway (PPP), generating NADPH to further inhibit ferroptosis. (Abbreviations: OXPHOS, oxidative phosphorylation; FAD, flavin adenine dinucleotide; IMM, inner mitochondrial membrane; LDHA, lactate dehydrogenase A; MCT4, monocarboxylate transporter 4.) Created in <a href="http://BioRender.com" target="_blank">BioRender.com</a> (accessed on 9 September 2024).</p>
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<p>Amino acid metabolism and ferroptosis: (<b>A</b>) The Xc-system, comprising SLC7A11 and SLC3A2, transports cystine and glutamate (Glu). Cystine undergoes a series of enzymatic reactions to form glutathione (GSH), which, under the catalysis of glutathione peroxidase 4 (GPX4), scavenges lipid peroxides to inhibit ferroptosis. Inhibition of the Xc-system impairs GSH formation, thereby promoting ferroptosis. (<b>B</b>) Glutamine (Gln) is catalyzed by Glutaminase 2 (GLS2) to produce glutamate, which is further catalyzed to form α-ketoglutarate, entering the tricarboxylic acid cycle (TCA) and generating ROS that promote ferroptosis. (Abbreviations: GCL, glutamate cysteine ligase; GS, glutathione synthetase; SLC1A5, solute carrier family 1 member 5; GDH, glutamate dehydrogenase; GOT, glutamic oxaloacetic transaminase.) Created in <a href="http://BioRender.com" target="_blank">BioRender.com</a> (accessed on 9 September 2024).</p>
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<p>Lipid metabolism and ferroptosis: Lipid peroxides directly induce the occurrence of ferroptosis. The formation of lipid peroxides proceeds via two pathways: enzymatic and non-enzymatic reactions. (<b>A</b>) Enzymatic reaction: AA-PL in the cell membrane is catalyzed by acyl-CoA synthetase long-chain family member 4 (ACSL4) to form AA-CoA, which is then sequentially catalyzed by lysophosphatidylcholine acyltransferase 3 (LPCAT3) and lipoxygenases (LOXs) to generate lipid peroxides. (<b>B</b>) Non-enzymatic reaction: Iron ions undergo the Fenton reaction to produce lipid peroxides. Created in <a href="http://BioRender.com" target="_blank">BioRender.com</a> (accessed on 9 September 2024).</p>
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14 pages, 1307 KiB  
Article
Effects of Marigold Extract and Carophyll Red on Growth, Body Color Development, Antioxidant Properties, and Innate Immunity in the Ornamental Fish Golden Severum (Heros efasciatus)
by Jung-Jin Park, Jun-Sung Bae, Chae-Won Lee, Chan-Young Yang, Eun-Ha Jeong, Kwan-Ha Park, Jung-Soo Seo, Mun-Gyeong Kwon and Ji-Hoon Lee
Life 2024, 14(12), 1660; https://doi.org/10.3390/life14121660 - 13 Dec 2024
Viewed by 327
Abstract
The body color state is an important determinant of the value of golden severum (Heros efasciatus)—a popular ornamental fish. The use of dietary supplements to improve the color development and health of this species is unexplored. Herein, the effects of marigold [...] Read more.
The body color state is an important determinant of the value of golden severum (Heros efasciatus)—a popular ornamental fish. The use of dietary supplements to improve the color development and health of this species is unexplored. Herein, the effects of marigold extract (MG) and carophyll red (CR) are examined on the growth, body color development, antioxidant properties, and innate immunity in golden severum. Fish were maintained under controlled water quality conditions (pH, temperature, and dissolved oxygen) and fed six experimental diets containing either 0% MG and CR, 1% MG, 2% MG, 5% MG, 0.5% CR, or 2% CR for five weeks. Both MG and CR significantly decreased lipid peroxide levels in hepatic tissues. In contrast, only MG enhanced the activities of reactive oxygen species (ROS)-scavenging enzymes (superoxide dismutase and catalase). Although MG and CR decreased the respiratory burst activity of splenic leukocytes, other innate immune parameters remained unchanged. Additionally, MG and CR stimulated body color development patterns in golden severum that reflect their unique coloring principles. The ROS-scavenging abilities of MG and CR appear to be related to their antioxidant activity. Hence, MG and CR at the optimal levels of 1.0% and 0.5%, respectively, can improve the body color of golden severum and protect against oxidative stress. Full article
(This article belongs to the Special Issue Feature Papers in Animal Science: 3rd Edition)
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<p>Approximate measurement points of chromaticity (body color) on a typical golden severum fed a 1.0% MG diet for five weeks.</p>
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<p>Effects of 5-week MG and CR feeding on body color values of golden severum. Data are expressed as the mean ± standard deviation from 60 fish (triplicate tanks of 20 fish each). * Significant difference from control at <span class="html-italic">p</span> &lt; 0.05 with Dunnett’s analysis. Con represents the control group, and the numbers on the <span class="html-italic">X</span>-axis indicate the diet percentage. (<b>a</b>) Lightness value (L), representing the brightness of the body color, measured using a chromatometer; (<b>b</b>) Red–green value (a), indicating the red or green intensity of the body color; (<b>c</b>) Yellow–blue value (b), reflecting the yellow or blue intensity of the body color. Values were expressed using Hunter chromaticity scales, with background reference values for the mounting board set to L = 101.42, a = 0.08, and b = −0.32.</p>
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<p>Effects of 5-week MG and CR feeding on hepatic antioxidant parameters of golden severum. Data are expressed as the mean ± standard deviation from 12 fish per group (4 fish from each of the 3 tanks). * Significant difference from control at <span class="html-italic">p</span> &lt; 0.05 with Dunnett’s analysis. Con, control; numbers on the <span class="html-italic">X</span>-axis denote diet percentage. (<b>a</b>) SOD activity (inhibition rate, %), reflecting the superoxide dismutase-mediated inhibition of WST-1 formazan formation; (<b>b</b>) GST activity (nmol/min/mg protein), based on glutathione conjugation with CDNB; (<b>c</b>) Lipid peroxide level (MDA, nmol/mg protein), indicating malondialdehyde levels; (<b>d</b>) CAT activity (nmol/min/mg protein), measured via formaldehyde formation.</p>
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<p>Effects of 5-week MG and CR feeding on direct and indirect innate immune parameters of golden severum. Data are expressed as the mean ± standard deviation from 12 fish per group (4 fish from each of the 3 tanks). * Significant difference from control at <span class="html-italic">p</span> &lt; 0.05 with Dunnett’s analysis. Con, control; numbers on the <span class="html-italic">X</span>-axis denote diet percentage. (<b>a</b>) Lysozyme activity (U/mL): measured by the degradation rate of <span class="html-italic">Micrococcus lysodeikticus</span> cell walls; (<b>b</b>) Respiratory burst activity (A630 nm): assessed by ROS production using PMA and nitroblue tetrazolium; (<b>c</b>) Phagocytic rate (%): percentage of splenic leukocytes engulfing zymosan particles; (<b>d</b>) Phagocytic index: average number of zymosan particles engulfed per leukocyte.</p>
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21 pages, 8441 KiB  
Article
Effects of Malondialdehyde on Growth Performance, Gastrointestinal Health, and Muscle Quality of Striped Catfish (Pangasianodon hypophthalmus)
by Cong Peng, Xinlangji Fu, Yumeng Zhang, Haitao Zhang, Yuantu Ye, Junming Deng and Beiping Tan
Antioxidants 2024, 13(12), 1524; https://doi.org/10.3390/antiox13121524 - 13 Dec 2024
Viewed by 381
Abstract
Malondialdehyde (MDA) is a reactive carbonyl compound produced through lipid peroxidation during feed storage, which poses a significant threat to fish health. This study aimed to evaluate the effects of dietary MDA on the growth rate, gastrointestinal health, and muscle quality of striped [...] Read more.
Malondialdehyde (MDA) is a reactive carbonyl compound produced through lipid peroxidation during feed storage, which poses a significant threat to fish health. This study aimed to evaluate the effects of dietary MDA on the growth rate, gastrointestinal health, and muscle quality of striped catfish (Pangasianodon hypophthalmus). A basal diet (M0) containing 34% crude protein and 10.5% crude lipid was formulated. Each group was sprayed with malondialdehyde solution (0, 5, 10, 20, 40, and 80 mg/kg, on dietary crude lipid basis; 0, 0.53, 1.07, 2.13, 4.26, and 8.52 mg/kg, on dietary basis) before feeding, respectively. Each diet was randomly assigned to triplicates of 30 striped catfish (initial weight 31.38 g) per net cage. After 8 weeks, dietary inclusion of MDA regardless of level significantly depressed the growth rate and feed utilization. The extent of structural damage to the gastrointestinal tract increased progressively with increasing dietary MDA levels. The extent of damage to the intestinal biological barrier (intestinal microbial structure), chemical barrier (trypsin, lipase, amylase, and maltase activity), physical barrier (zonula occludent-2, occludin, claudin 7α, and claudin 12 relative expression), and immune barrier (contents of complement 4, complement 3, immunoglobulin M, and lysozyme activity) was dose-related to dietary MDA. Moreover, a linear decline in the activities of intestinal antioxidant enzymes (catalas, superoxide dismutase, et al.) and anti-inflammatory factor (transforming growth factor beta1, interleukin 10) relative expression was noted alongside an increase in dietary MDA content. In contrast, the relative expression levels of intestinal inflammatory factor (interleukin 8, transcription factor p65, tumor necrosis factor alpha) relative expression displayed an opposing trend. Additionally, dietary MDA exerted a linear influence on muscle color and texture characteristics. In conclusion, high doses of MDA (5–80 mg/kg) reduced the growth performance of striped catfish, attributed to linear damage to the gastrointestinal tract, a linear decrease in antioxidant function, and the occurrence of an inflammatory response. High doses of MDA (>40 mg/kg) were observed to significantly increase dorsal muscle b-value and induce muscle yellowing. Full article
(This article belongs to the Special Issue Oxidative Stress and Nutrition in Aquatic Animals)
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<p>The scanning electron microscope of the stomach in striped catfish-fed diets with various levels of malondialdehyde (×3000). (<b>A</b>) Diet M0, (<b>B</b>) Diet M20, (<b>C</b>) Diet M80. Striped catfish-fed the M0 diet (<b>A</b>) showed the stomach mucosal surfaces were smooth, soft, and free of erosions, while stomach mucosal cells were ruptured, and the mucosa was extensively ulcerated in fish fed the M20 (<b>B</b>) and M80 (<b>C</b>) diets.</p>
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<p>The stomach histomorphology of striped catfish-fed diets with various levels of malondialdehyde (H&amp;E staining, ×40). (<b>A</b>) Diet M0, (<b>B</b>) Diet M5, (<b>C</b>) Diet M10, (<b>D</b>) Diet M20, (<b>E</b>) Diet M40, (<b>F</b>) Diet M80. VH, villi height; VW, villi width; MT, muscular layer thickness. Striped catfish-fed the M0 (<b>A</b>) and M5 (<b>B</b>) diets exhibited healthy stomach structure with intact columnar epithelium, stomach glands, mucosa, and submucosa, while damaged stomach tissue with the degenerated columnar epithelium, atrophied stomach glands, and destructed villus integrity was observed in striped catfish fed the M10 (<b>C</b>), M20 (<b>D</b>), M40 (<b>E</b>), and M80 (<b>F</b>) diets.</p>
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<p>The stomach histomorphology of striped catfish-fed diets with various levels of malondialdehyde (H&amp;E staining, ×40). (<b>A</b>) Diet M0, (<b>B</b>) Diet M5, (<b>C</b>) Diet M10, (<b>D</b>) Diet M20, (<b>E</b>) Diet M40, (<b>F</b>) Diet M80. VH, villi height; VW, villi width; MT, muscular layer thickness. Striped catfish-fed the M0 (<b>A</b>) and M5 (<b>B</b>) diets exhibited healthy stomach structure with intact columnar epithelium, stomach glands, mucosa, and submucosa, while damaged stomach tissue with the degenerated columnar epithelium, atrophied stomach glands, and destructed villus integrity was observed in striped catfish fed the M10 (<b>C</b>), M20 (<b>D</b>), M40 (<b>E</b>), and M80 (<b>F</b>) diets.</p>
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<p>The intestinal histomorphology of striped catfish-fed diets with various levels of malondialdehyde (H&amp;E staining, ×100). (<b>A</b>) Diet M0, (<b>B</b>) Diet M5, (<b>C</b>) Diet M10, (<b>D</b>) Diet M20, (<b>E</b>) Diet M40, (<b>F</b>) Diet M80. VH, villi height; VW, villi width; MT, muscular layer thickness. Striped catfish-fed the M0 (<b>A</b>) and M5 (<b>B</b>) diets exhibited normal intestines with intact villus, while the damaged intestine with shortened villus and thinned lamina propria was observed in fish fed the M10 (<b>C</b>), M20 (<b>D</b>), M40 (<b>E</b>), and M80 (<b>F</b>) diets.</p>
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<p>The intestinal microbial diversity of striped catfish-fed diets containing different levels of malondialdehyde. (<b>A</b>) Venn diagram based on the OTU level (each group is represented by a different color. The intersection part of the figure represents the common OTUs between different groups), (<b>B</b>) Principal Co-ordinates analysis diagram based on the OTU level and weighted_unifrac Distance (The dots represent a group, PCoA1 represents the principal coordinate component that best explains the variation in the data, and PCoA2 represents the principal coordinate component that accounts for most of the remaining variation).</p>
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<p>The intestinal microbial composition of striped catfish-fed diets containing different malondialdehyde levels. (<b>A</b>) Plot of microbial composition based on the phylum level; (<b>B</b>) Plot of microbial composition based on the genus level; (<b>C</b>) Heat map of microbial abundance based on the phylum level; (<b>D</b>) Heat map of microbial abundance based on the genus level. (<b>C</b>,<b>D</b>) each column represents a group; each row represents a species. The colors represent species abundance; nearer to orange is less abundance and nearer to blue is more abundance. (<b>E</b>) Evolutionary clade chart, (<b>F</b>) LDA distribution histogram (<b>E</b>) Illustrating the differential species across various taxonomic ranks, from inner to outer in the following sequence: Phylum, Class, Order, Family, Genus, Species. (<b>F</b>) bacterial taxa differentially represented in the intestinal microbial populations of different groups were identified by LEfSe using an LDA, with LDA scores &gt; 3 and <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The intestinal microbial functional prediction at the species level in striped catfish-fed diets with various malondialdehyde levels. (<b>A</b>) PICRUSt2 heat map of third-level functional pathway annotation (each column represents a group; each row represents a functional pathway. The colors represent the relative abundance of the pathway; the warmer the color (closer to orange), the less the abundance; the cooler the color (closer to blue), the more the abundance). (<b>B</b>) PICRUSt2 Welch’s t-test of third-level functional pathway annotation (significant when <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>The scanning electron microscope of the intestine in striped catfish-fed diets with various levels of malondialdehyde (×7000). (<b>A</b>) Diet M0; (<b>B</b>) Diet M20; (<b>C</b>) Diet M80. MV, microvilli; M, mitochondria; TJ, tight junction; L, lysosome. Striped catfish-fed the M0 diet (<b>A</b>) showed normal enterocytes, while enterocytes with sparse and disorganized microvilli, swollen mitochondria, and widened intercellular space were observed in fish fed the M20 (<b>B</b>) and M80 (<b>C</b>) diets.</p>
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<p>The relative expression of inflammatory response and tight junction protein-related genes in striped catfish-fed diets with various levels of malondialdehyde (<span class="html-italic">n</span> = 6). Values are presented as means of triplication; means in the same row with different superscript letters represented a significant difference (<span class="html-italic">p</span> &lt; 0.05); PSE = pooled standard error of means. <span class="html-italic">ZO-2</span>, zonula occludent 2; <span class="html-italic">tnf-α</span>, tumor necrosis factor alpha; <span class="html-italic">p65</span>, transcription factor p65; <span class="html-italic">il-8</span>, interleukin 8; <span class="html-italic">il-10</span>, interleukin 10; <span class="html-italic">tgf-β1</span>, transforming growth factor beta1. Linear regression equation (where y is the response and x is the level of malondialdehyde in diet), R<sup>2</sup>, and <span class="html-italic">p</span>-Value (significant when <span class="html-italic">p</span> &lt; 0.05) are also given.</p>
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20 pages, 6210 KiB  
Article
Clinicopathological Studies on the Impact of Grape Seed Extract and L-Carnitine as Cardioprotective Agents Against Doxorubicin-Induced Toxicity in Rats
by Tahany Saleh Aldayel, Omnia E. Kilany, Heba Nageh Gad El-Hak, Heba M. A. Abdelrazek, Osama Abdallah and Donia E. Omar
Life 2024, 14(12), 1656; https://doi.org/10.3390/life14121656 - 13 Dec 2024
Viewed by 414
Abstract
Doxorubicin (DOX) cancer therapy induces serious cardiotoxicity as a side effect. This study aimed to investigate the cardioprotective effects of grape seed extract (GSE) and L-Carnitine (L-CA) against DOX-induced cardiac toxicity in male rats. Six groups of male albino rats were used: G1 [...] Read more.
Doxorubicin (DOX) cancer therapy induces serious cardiotoxicity as a side effect. This study aimed to investigate the cardioprotective effects of grape seed extract (GSE) and L-Carnitine (L-CA) against DOX-induced cardiac toxicity in male rats. Six groups of male albino rats were used: G1 (control); G2 (GSE), given grape seed extract (100 mg/kg b.wt.) orally for 35 days; G3 (L-CA) (150 mg/kg b.wt.); Group 4 (DOX-induced cardiotoxicity), given DOX (10 mg/kg b.wt., i.p.) on the 28th day of the experiment; G5 (GSE + DOX), given GSE and DOX as previously mentioned; and G6 (L-CA + DOX), given L-CA and DOX as previously mentioned. Electrocardiographic evaluation, lipid profile, lipid peroxidation and antioxidants, serum cardiac markers, and inflammatory markers were estimated. Histopathological evaluation of cardiac tissue was also examined. Key findings showed that DOX induced ECG abnormalities lipid peroxidation, reduced antioxidants, and elevated cardiac and inflammatory markers. GSE and L-CA significantly ameliorated ECG abnormalities, reduced lipid peroxidation, improved antioxidant enzymes and serum cardiac markers, and reduced inflammation. These findings suggest that GSE and L-CA exhibit substantial cardioprotective effects in DOX-induced cardiotoxicity via their antioxidant and anti-inflammatory potentials. Full article
(This article belongs to the Special Issue Advancements in Heart Failure Research)
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Figure 1
<p>Phytochemical analysis of grape seed extract.</p>
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<p>(<b>A</b>) Representative ECG on day 29 of the experiment. (<b>B</b>) Representative ECG on day 35 of the experiment. Group GSE: grape seed extract, Group L-CA: L-Carnitine; Group DOX: Doxorubicin; Group GSE + DOX: Grape seed extract with Doxorubicin, Group L-CA+ DOX: L-Carnitine with Doxorubicin. Labels a, b, and c denote the following ECG parameters: (a) ST-segment elevation, (b) QRS interval and (c) QT interval.</p>
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<p>The impact of doxorubicin, grape seed extract, and L-carnitine on heart rate (HR), QRS complex, QT interval, and ST segment amplitude. (<b>A</b>) Representation of the changes in ECG on day 29 of the experiment (<span class="html-italic">n</span> = 5). Data for each rat were obtained from 4 readings/rat. (<b>B</b>) Representation of the changes in ECG on day 35 of the experiment (<span class="html-italic">n</span> = 5). Data for each rat were obtained from 4 readings/rat.</p>
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<p>Photomicrographs of cardiac myocytes from the control, GSE, and L-CA groups (<b>a</b>–<b>c</b>) on day 29 of the experiment displayed normal cardiac myofibrillar structure (MF) with striations, branched appearance, central nucleus, and continuity with adjacent myofibrils. In contrast, the DOX group (<b>d</b>) revealed several abnormalities, including nuclear pyknosis and leukocyte infiltration (indicated by arrows). The DOX + GSE and DOX + L-CA groups (<b>e</b>,<b>f</b>) showed a restoration to normal cardiac myofibrillar structure (C) with clear striations. (H&amp;E, 400×).</p>
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<p>Photomicrographs of cardiac myocytes from the control, GSE, and L-CA groups (<b>a</b>–<b>c</b>) on day 35 of the experiment showed normal cardiac myofibrillar structure (MF) with striations, a branched appearance, a central nucleus, and continuity with adjacent myofibrils. In contrast, the DOX group (<b>d</b>) exhibited signs of myocardial infarction, including focal myolysis, non-visible nuclei, and darker, irregular necrotic contraction bands (indicated by arrowheads). The DOX + GSE and DOX + L-CA groups (<b>e</b>,<b>f</b>) displayed a return to normal cardiac myofibrillar structure (C) with clear striations. (H&amp;E, 400×).</p>
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<p>Photomicrographs of Masson-trichrome-stained cardiac myocytes of low and high magnification from the control (<b>a</b>,<b>g</b>), GSE (<b>b</b>,<b>h</b>), and L-CA groups (<b>c</b>,<b>i</b>) on day 35 of the experiment depicted normal cardiac myofibrillar structure with striations, a branched appearance, a central nucleus, and continuity with adjacent myofibrils. In contrast, the DOX group (<b>d</b>,<b>j</b>) exhibited interstitial fibrosis, with an increase in the percentage area of collagen fiber (arrow). However, the GSE + DOX (<b>e</b>,<b>k</b>) and L-CA + DOX groups (<b>f</b>,<b>l</b>) displayed a restoration of normal cardiac myofibrillar structure with clear striations, accompanied by a decrease in the percentage area of collagen fiber. (100× &amp; 400×). The mean percentage area of collagen fiber (<b>m</b>) was quantified across (<span class="html-italic">n</span> = 5) rats/group. Each set of rat data was obtained from 12 measurements. Group GSE: grape seed extract; Group L-CA: L-Carnitine; Group DOX: Doxorubicin; Group GSE + DOX: Grape seed extract with Doxorubicin; Group L-CA + DOX: L-Carnitine with Doxorubicin.</p>
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14 pages, 2155 KiB  
Article
Integration of Copper Toxicity Mechanisms in Raphidocelis subcapitata: Advancing Insights at Environmentally Relevant Concentrations
by Manuela D. Machado and Eduardo V. Soares
Toxics 2024, 12(12), 905; https://doi.org/10.3390/toxics12120905 - 13 Dec 2024
Viewed by 409
Abstract
This work aimed to characterize the impact of copper (Cu), at environmentally relevant concentrations, using the freshwater microalga Raphidocelis subcapitata. Algae were incubated with 33 or 53 µg/L Cu, in OECD medium, and toxic impacts were evaluated over 72 h, using different [...] Read more.
This work aimed to characterize the impact of copper (Cu), at environmentally relevant concentrations, using the freshwater microalga Raphidocelis subcapitata. Algae were incubated with 33 or 53 µg/L Cu, in OECD medium, and toxic impacts were evaluated over 72 h, using different cellular and biochemical biomarkers. The exposure to 33 µg/L Cu had an algistatic effect: slowing growth and reducing algal population (53%, at 72 h) without compromising the cell membrane. This Cu concentration promoted a transient reduction in chlorophyll a (chla) content and typical markers of oxidative stress: increased levels of reactive oxygen species (ROS), augmented catalase (CAT) activity, and lipid peroxidation (malondialdehyde, MDA). Algae exposed to 53 µg/L Cu, suffered a severe effect with a 93% reduction in the number of cells, 50% decrease in chla content, and diminished (17%) maximum photochemical quantum yield of PSII (Fv/Fm). This population also presented increased levels of ROS and MDA, 33 and 20 times higher than the control, respectively, at 72 h, augmented CAT activity, and permeabilized cell membrane (5%, at 72 h). These findings provide valuable insights into Cu toxicity in aquatic ecosystems, highlighting the biochemical and physiological impacts at environmentally relevant concentrations. Full article
(This article belongs to the Section Ecotoxicology)
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<p>Effect of Cu on <span class="html-italic">R. subcapitata</span> proliferation capacity. Evolution of algal population on OECD medium in the absence (control) or the presence of Cu. Data are presented as mean values ± SD (error bars). At each time, the statistical difference between control and Cu-treated cells was tested using unpaired Student’s <span class="html-italic">t</span>-test; the means with (*) are significantly different from the control (<span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">n</span> = 6).</p>
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<p>Impact of Cu on <span class="html-italic">R. subcapitata</span> chlorophyll <span class="html-italic">a</span> content. Data are presented as mean values ± SD; mean values with different letters are significantly different (<span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">n</span> = 5, Kruskal–Wallis test, Dunn’s post hoc test).</p>
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<p>Influence of Cu on <span class="html-italic">R. subcapitata</span> photosynthetic activity evaluated by pulse-amplitude-modulation fluorescence assay. (<b>A</b>) Maximum photochemical quantum yield of PSII (<span class="html-italic">F</span><sub>v</sub>/<span class="html-italic">F</span><sub>m</sub>). (<b>B</b>) Effective photochemical quantum yield of PSII (Φ<sub>PSII</sub>). (<b>C</b>) Electron transport rate (<span class="html-italic">ETR</span>). (<b>D</b>) Non-photochemical quenching (<span class="html-italic">NPQ</span>). Data are presented as mean values ± SD. At each time, the statistical difference between control and Cu-treated cells was tested using unpaired Student’s <span class="html-italic">t</span>-test; the means with (*) are significantly different from the control (<span class="html-italic">p</span> &lt; 0.05, <span class="html-italic">n</span> = 5).</p>
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<p>ROS accumulation and antioxidant activity of <span class="html-italic">R. subcapitata</span> cells exposed to Cu. (<b>A</b>) Reactive oxygen species (ROS) production. (<b>B</b>) Visualization of the intracellular accumulation of ROS (green cells) by epifluorescence microscopy and using the H<sub>2</sub>DCFDA probe on algae not exposed or exposed to Cu for 72 h. (<b>C</b>) Carotenoids content. (<b>D</b>) Superoxide dismutase (SOD) activity. (<b>E</b>) Catalase (CAT) activity. Data are presented as mean values ± SD. At each time, the statistical difference between control and Cu-treated cells was tested using unpaired Student’s <span class="html-italic">t</span>-test; the means with (*) are significantly different from the control (<span class="html-italic">p</span> &lt; 0.05; <span class="html-italic">n</span> = 4).</p>
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<p>Lipid peroxidation and loss of cell membrane integrity of <span class="html-italic">R. subcapitata</span> exposed to Cu. (<b>A</b>) Malondialdehyde (MDA) content (lipid peroxidation). (<b>B</b>) Loss of plasma membrane integrity. Data are presented as mean values ± SD. At each time, the statistical difference between control and Cu-treated cells was tested using unpaired Student’s <span class="html-italic">t</span>-test; the means with (*) are significantly different from the control (<span class="html-italic">p</span> &lt; 0.05; <span class="html-italic">n</span> = 3 for lipid peroxidation and <span class="html-italic">n</span> = 5 for cell membrane integrity).</p>
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<p>Proposal of the mechanism of action (toxicity pathway) of Cu, at environmentally relevant concentrations on the freshwater alga <span class="html-italic">R. subcapitata</span> based on the results here presented and previous works [<a href="#B13-toxics-12-00905" class="html-bibr">13</a>,<a href="#B23-toxics-12-00905" class="html-bibr">23</a>,<a href="#B27-toxics-12-00905" class="html-bibr">27</a>]. Variations in cellular responses are symbolized by “+” or “−“ for an increase or decrease, respectively. CAT—catalase, GSH—reduced glutathione, ROS—reactive oxygen species.</p>
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