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18 pages, 4334 KiB

Open AccessArticle

Phytochemical Analysis and Multifaceted Biomedical Activities of Nitraria retusa Extract as Natural Product-Based Therapies

by Manal M. Khowdiary, Zinab Alatawi, Amirah Alhowiti, Mohamed A. Amin, Hussam Daghistani, Faisal Miqad K. Albaqami, Mohamed Ali Abdel-Rahman, Ahmed Ghareeb, Nehad A. Shaer, Ahmed M. Shawky and Amr Fouda

Life 2024, 14(12), 1629; https://doi.org/10.3390/life14121629 - 9 Dec 2024

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Abstract

This study examined the phytochemical profile and biomedical activities of Nitraria retusa, a halophytic and drought-resistant shrub. HPLC analysis showed gallic acid (1905.1 μg/g), catechin (1984.1 μg/g), and ellagic acid (2671.1 μg/g) as the primary constituents, while FT-IR analysis revealed a complex [...] Read more.

This study examined the phytochemical profile and biomedical activities of Nitraria retusa, a halophytic and drought-resistant shrub. HPLC analysis showed gallic acid (1905.1 μg/g), catechin (1984.1 μg/g), and ellagic acid (2671.1 μg/g) as the primary constituents, while FT-IR analysis revealed a complex organic profile with significant functional groups. The extract demonstrated strong antioxidant activity in DPPH assays, outperforming ascorbic acid (IC₅₀ = 18.7 ± 1.0 μg/mL) with an IC₅₀ of 16.4 ± 4.4 μg/mL. It demonstrated specific antiproliferative effects on cancer cell lines as it showed selective cytotoxicity against cancer cell lines; normal WI38 cells were largely unaffected, showing 50.0% viability at 125 μg/mL. The most sensitive cell line was Caco2, which showed 50.0% viability at 125 μg/mL. Anti-diabetic properties were exhibited by means of inhibition of α-amylase (IC₅₀ = 68.2 ± 4.2 μg/mL) and α-glucosidase (IC₅₀ = 22.8 ± 3.3 μg/mL). Additionally, antimicrobial activity was observed to be broad-spectrum, and it was most effective against E. coli (32.6 mm inhibition zone at 400 μg/mL) and Penicillium glabrum (35.3 mm at 400 μg/mL). These findings highlight the potential of N. retusa in developing plant-based therapeutic approaches. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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N. retusa plant collected from Wadi Hagul, Eastern Desert, Egypt. Full article ">Figure 2
HPLC standards used in the current investigation. Full article ">Figure 3
Phytochemical fingerprint of Nitraria retusa extract based on HPLC analysis. Full article ">Figure 4
DPPH radical scavenging activity of Nitraria retusa extract vs. ascorbic acid (1.95–1000 μg/mL, n = 3). Different letters (a and b) on the bars indicate significant differences between treatments at the same concentration. *** p < 0.001 indicate significant differences from control group (p < 0.05, p < 0.01, and p < 0.001, respectively); (n = 3). Full article ">Figure 5
Dose-dependent cytotoxicity of plant extract on Wi38, Caco2, and Mcf7 cell lines. Data represents mean cell viability (%). Different letters (a, b, and c) on the bars at the same concentration indicate significant differences. *, **, and *** p < 0.001 indicate significant differences from control group (p < 0.05, p < 0.01, and p < 0.001, respectively); (n = 3). Full article ">Figure 6
Dose-dependent α-amylase inhibition by Nitraria retusa extract vs. acarbose (1.95–1000 μg/mL). Different letters (a and b) on the bars at the same concentration indicate significant differences. *** p < 0.001 indicate significant differences from control group (p < 0.05, p < 0.01, and p < 0.001, respectively); (n = 3). Full article ">Figure 7
Profiles of glucosidase inhibition by plant extract and acarbose (1.95–1000 μg/mL. Different letters (a and b) on the bars at the same concentration indicate significant differences. *** p < 0.001 indicate significant differences from control group (p < 0.05, p < 0.01, and p < 0.001, respectively); (n = 3). Full article ">Figure 8
Antimicrobial potency represented as the inhibition zone (mm) of plant extract and control against S. aureus, E. coli, C. albicans, and P. glabrum across concentrations (12.5–400 μg/mL). Different letters (a and b) on the bars at the same concentration indicate significant differences. **, and *** p < 0.001 indicate significant differences from control group (p < 0.05, p < 0.01, and p < 0.001, respectively); (n = 3). Full article ">

29 pages, 6570 KiB

Open AccessArticle

Clitoria ternatea L. (Butterfly Pea) Flower Against Endometrial Pain: Integrating Preliminary In Vivo and In Vitro Experimentations Supported by Network Pharmacology, Molecular Docking, and Molecular Dynamics Simulation Studies

by Najneen Ahmed, Nazifa Tabassum, Parisa Tamannur Rashid, Basrat Jahan Deea, Fahmida Tasnim Richi, Anshuman Chandra, Shilpi Agarwal, Saima Mollick, Kaushik Zaman Dipto, Sadia Afrin Mim and Safaet Alam

Life 2024, 14(11), 1473; https://doi.org/10.3390/life14111473 - 13 Nov 2024

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Abstract

Clitoria ternatea L. (CT) is a perennial herbaceous plant with deep blue flowers native to tropical Asia. This work explores the endometrial pain (EP) regulation of CT flower through a multifaceted approach. Phytochemical screening unveiled the presence of alkaloids, steroids, flavonoids, glycosides, and [...] Read more.

Clitoria ternatea L. (CT) is a perennial herbaceous plant with deep blue flowers native to tropical Asia. This work explores the endometrial pain (EP) regulation of CT flower through a multifaceted approach. Phytochemical screening unveiled the presence of alkaloids, steroids, flavonoids, glycosides, and tannins in CT flower methanolic extract (ME). In the in vitro membrane stabilizing experiment, the ME demonstrated 91.47% suppression of heat-induced hemolysis. Upon carrageenan-induced paw edema assay conducted on male Swiss albino mice at doses of 200 mg/kg and 400 mg/kg, 65.28% and 81.89% inhibition rates, respectively, of paw edema were reported. For the same doses, upon acetic acid-induced-writhing assay, 75.6% and 76.78% inhibition rates, respectively, were observed. For network pharmacology analyses, a protein–protein interaction network was constructed for 92 overlapping gene targets of CT and EP, followed by GO and KEGG pathway enrichment analyses. Network pharmacology-based investigation identified the anti-EP activity of CT to be mostly regulated by the proteins SRC homology, ESR1, and PI3KR1. Physicochemical, pharmacokinetic, and toxicity property predictions for the compounds with stable ligand–target interactions and a molecular dynamics simulation for the highest interacting complex further validated these findings. This work affirmed the anti-EP role of CT flower against EP, suggesting a probable molecular mechanism involved. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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23 pages, 2602 KiB

Open AccessArticle

Assessing the Optimal Antibacterial Action of Lavandula stoechas L., Thymus zygis L., and Eucalyptus camaldulensis Dehnh Essential Oils

by Farah Aabouch, Saoussan Annemer, Badr Satrani, Ismail Ettaleb, Mohammed Kara, Mohamed Ghanmi, Abdelaaty Abdelaziz Shahat, Ravish Choudhary, Abdellah Farah, Mohamed Ouajdi and Jamila Dahmani

Life 2024, 14(11), 1424; https://doi.org/10.3390/life14111424 - 5 Nov 2024

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Abstract

The use of combined essential oils (EOs) is a new technique that can improve their preservative effects while minimizing their sensory impact in foods. The aim of this study was to determine the chemical profile of three essential oils (EOs) extracted from Lavandula [...] Read more.

The use of combined essential oils (EOs) is a new technique that can improve their preservative effects while minimizing their sensory impact in foods. The aim of this study was to determine the chemical profile of three essential oils (EOs) extracted from Lavandula stoechas L. (Ls), Thymus zygis L. (Tz), and Eucalyptus camaldulensis Dehnh (Ec) and to evaluate their synergistic antibacterial activity for optimal inhibition against Bacillus subtilis, Escherichia coli, and Staphylococcus aureus using an augmented Simplex centroid mixing scheme. The essential oils were extracted by hydrodistillation and analyzed via gas chromatography–mass spectrometry. Anti-bacterial potency was evaluated by disk diffusion. Chemical analysis revealed the main compounds in Lavandula stoechas (Ls) essential oil: camphor (36.15%), followed by fenchone (16.57%) and Z-8-hydroxy linalool (8.28%). The Thymus zygis (Tz) essential oil is dominated by δ-terpineol (27.64%), δ-3-carene (15.7%), and thymol (14.17%). In contrast, the Eucalyptus camaldulensis (Ec) essential oil contains mainly 1,8-cineole (43.61%), γ-terpinene (11.71%), and α-terpineol (10.58%). The optimal mixture is the binary association of 40% E. camaldulensis EO and 60% T. zygis EO, which provides an effective inhibition diameter (ID) of 13.37 mm to inhibit S. aureus. Furthermore, the formulation of 27% and 73% EOs of E. camaldulensis and T. zygis, respectively, corresponds to the mixture required to achieve the optimum inhibition diameter (ID = 11.55 mm) against E. coli. In addition, the mixture of 29% EO of E. camaldulensis and 71% EO of T. zygis is the optimum mixture to inhibit B. subtilis, with an inhibition diameter of 12.31 mm. These findings highlight the potency of antibacterial formulations of these essential oils and suggest that they might be used as substitutes for conventional drugs to prevent the development of bacteria responsible for serious infections and food spoilage. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Positions of experimental points for augmented Simplex-centroid designs. Full article ">Figure 2
Average yield of the essential oils as a function the plants studied. Full article ">Figure 3
Scatter plot of observed values vs. predicted values. Full article ">Figure 4
(a) Two-dimensional mixing profile and (b) three-dimensional profile showing the zone of maximum di response based on the three constituents. Full article ">Figure 4 Cont.
(a) Two-dimensional mixing profile and (b) three-dimensional profile showing the zone of maximum di response based on the three constituents. Full article ">Figure 5
Desirability graph revealing the precise proportions of T. zygis, L. stoechas and E. camaldulensis EOs leading to the best antibacterial against S. aureus (a), E. coli (b) and B. subtilis (c). Full article ">Figure 5 Cont.
Desirability graph revealing the precise proportions of T. zygis, L. stoechas and E. camaldulensis EOs leading to the best antibacterial against S. aureus (a), E. coli (b) and B. subtilis (c). Full article ">

16 pages, 2953 KiB

Open AccessArticle

Effects of Drying Methods on the Phytochemical Contents, Antioxidant Properties, and Anti-Diabetic Activity of Nasturtium officinale R.Br. (Betong Watercress) from Southern Thailand

by Praporn Kijkuokool, Irina Stepanov, Sakaewan Ounjaijean, Pimpisid Koonyosying, Kittipan Rerkasem, Hataichanok Chuljerm, Wason Parklak and Kanokwan Kulprachakarn

Life 2024, 14(9), 1204; https://doi.org/10.3390/life14091204 - 23 Sep 2024

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Abstract

Nasturtium officinale R.Br. (Betong watercress) contains high levels of secondary metabolites that offer health benefits. However, fresh-cut watercress has a short shelf life. This study aimed to assess the effect of drying methods on the phytochemical contents, antioxidant activity, and anti-diabetic activity of [...] Read more.

Nasturtium officinale R.Br. (Betong watercress) contains high levels of secondary metabolites that offer health benefits. However, fresh-cut watercress has a short shelf life. This study aimed to assess the effect of drying methods on the phytochemical contents, antioxidant activity, and anti-diabetic activity of Betong watercress. The watercress was dried using three methods: roasting (R, 50 °C, 40 min); roasting and drying (RD, 40 min roasting at 50 °C and 1 h drying at 80 °C); and blanching, roasting, and drying (BRD, 30 s blanching at 80 °C, 20 min roasting at 50 °C, and 1 h drying at 80 °C). Aqueous extracts from each drying method were analyzed for total phenolic content, total flavonoid content, total glucosinolate content, antioxidant activities (FRAP, DPPH, and ABTS assays), and α-amylase enzyme inhibition. From the results, the R method provided the highest level of total phenolic, total flavonoid, and total glucosionolate content compared to the RD and BRD methods. Similarly, antioxidant activities and α-amylase enzyme inhibition were highest in the R method, followed by the RD and BRD methods. Our results demonstrate that roasting of Betong watercress without the addition of blanching or drying effectively preserves the phytochemical contents, antioxidant activities, and anti-diabetic activity. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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13 pages, 2091 KiB

Open AccessArticle

From Nature to Treatment: The Impact of Pterostilbene on Mitigating Retinal Ischemia–Reperfusion Damage by Reducing Oxidative Stress, Inflammation, and Apoptosis

by Beáta Pelles-Taskó, Réka Szekeres, Barbara Takács, Anna Szilágyi, Dóra Ujvárosy, Mariann Bombicz, Dániel Priksz, Balázs Varga, Rudolf Gesztelyi, Zoltán Szabó, Zoltán Szilvássy and Béla Juhász

Life 2024, 14(9), 1148; https://doi.org/10.3390/life14091148 - 11 Sep 2024

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Abstract

Retinal ischemia–reperfusion (I/R) injury is a critical pathogenic mechanism in various eye diseases, and an effective therapeutic strategy remains unresolved. Natural derivatives have recently reemerged; therefore, in our present study, we examined the potential therapeutic effects of a stilbenoid that is chemically related [...] Read more.

Retinal ischemia–reperfusion (I/R) injury is a critical pathogenic mechanism in various eye diseases, and an effective therapeutic strategy remains unresolved. Natural derivatives have recently reemerged; therefore, in our present study, we examined the potential therapeutic effects of a stilbenoid that is chemically related to resveratrol. Pterostilbene, recognized for its anti-inflammatory, anti-carcinogenic, anti-diabetic, and neuroprotective properties, counteracts oxidative stress during I/R injury through various mechanisms. This study explored pterostilbene as a retinoprotective agent. Male Sprague Dawley rats underwent retinal I/R injury and one-week reperfusion and were treated with either vehicle or pterostilbene. After this functional electroretinographical (ERG) measurement, Western blot and histological analyses were performed. Pterostilbene treatment significantly improved retinal function, as evidenced by increased b-wave amplitude on ERG. Histological studies showed reduced retinal thinning and preserved the retinal structure in the pterostilbene-treated groups. Moreover, Western blot analysis revealed a decreased expression of glial fibrillary acidic protein (GFAP) and heat shock protein 70 (HSP70), indicating reduced glial activation and cellular stress. Additionally, the expression of pro-apoptotic and inflammatory markers, poly(ADP-ribose) polymerase 1 (PARP1) and nuclear factor kappa B (NFκB) was significantly reduced in the pterostilbene-treated group. These findings suggest that pterostilbene offers protective effects on the retina by diminishing oxidative stress, inflammation, and apoptosis, thus preserving retinal function and structure following I/R injury. This study underscores pterostilbene’s potential as a neuroprotective therapeutic agent for treating retinal ischemic injury and related disorders. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Comparison of a-wave and b-wave responses in ERG: (A) amplitude analysis of a-wave responses across different experimental groups; (B) amplitude analysis of b-wave responses across different experimental groups. Data represent mean ± SEM; * p < 0.05. ns = no significant difference. Full article ">Figure 2
Statistically most significant comparisons in scotopic ERG measurements; flash intensity: mcd·s·m−2: (A) average b-wave amplitudes (µV) of the different groups at 1000 mcd·s·m−2 light intensity; (B) average b-wave amplitudes (µV) at 3000 mcd·s·m−2 light intensity; (C) average b-wave amplitudes (µV) at 25,000 mcd·s·m−2 light intensity (µV). All results are plotted as group average ± SEM. Statistically significant comparisons are denoted by * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Full article ">Figure 3
Implicit time analysis of a-wave and b-wave responses in electroretinography (ERG): (A) comparison of a-wave implicit times among different treatment groups; (B) comparison of b-wave implicit times among different treatment groups. The statistical analysis showed no significant differences in implicit times among the different treatment groups. Data represent mean ± SEM; p > 0.05. ns = no significant difference. Full article ">Figure 4
Expression levels of glial fibrillary acidic protein (GFAP) and heat shock protein 70 (HSP70) in retinal tissue: (A) Western blot analysis revealing a notable increase in GFAP expression in different groups; (B) Western blot analysis demonstrating a similar pattern with elevated levels of HSP70 in different groups. Data represent mean ± SEM; *** p < 0.001, **** p < 0.0001, n = 4 per group. Expression levels of poly(ADP-ribose) polymerase 1 (PARP1) and nuclear factor kappa B (NFkB) in retinal tissue; (C) Western blot analysis showing significantly elevated expression of PARP1 in different groups; (D) Western blot analysis revealing a similar pattern with significantly increased levels of NFkB in different groups. Data represent mean ± SEM; ** p < 0.01, **** p < 0.0001, n = 4 per group. Full article ">Figure 5
Microscopic analysis of rat retinal sections: (A) representative images of retinal thickness in the different treatment groups (from left to right): MUCI NO IR, MUCI IR, PTER NO IR, and PTER IR; (B) graphs showing statistical analysis results of histology sections of the different groups. Data represent mean ± SEM; **** p < 0.0001. Full article ">

19 pages, 2230 KiB

Open AccessArticle

Evaluation of the Interaction between Carvacrol and Thymol, Major Compounds of Ptychotis verticillata Essential Oil: Antioxidant, Anti-Inflammatory and Anticancer Activities against Breast Cancer Lines

by Mohamed Taibi, Amine Elbouzidi, Mounir Haddou, Abdellah Baraich, Douaae Ou-Yahia, Reda Bellaouchi, Ramzi A. Mothana, Hanan M. Al-Yousef, Abdeslam Asehraou, Mohamed Addi, Bouchra El Guerrouj and Khalid Chaabane

Life 2024, 14(8), 1037; https://doi.org/10.3390/life14081037 - 20 Aug 2024

Cited by 2 | Viewed by 1517

Abstract

The objective of this study was to evaluate the antioxidant, anti-inflammatory, and anticancer properties of thymol, carvacrol, and their equimolar mixture. Antioxidant activities were assessed using the DPPH, ABTS, and ORAC methods. The thymol/carvacrol mixture exhibited significant synergism, surpassing the individual compounds and [...] Read more.

The objective of this study was to evaluate the antioxidant, anti-inflammatory, and anticancer properties of thymol, carvacrol, and their equimolar mixture. Antioxidant activities were assessed using the DPPH, ABTS, and ORAC methods. The thymol/carvacrol mixture exhibited significant synergism, surpassing the individual compounds and ascorbic acid in DPPH (IC₅₀ = 43.82 ± 2.41 µg/mL) and ABTS (IC₅₀ = 23.29 ± 0.71 µg/mL) assays. Anti-inflammatory activity was evaluated by inhibiting the 5-LOX, COX-1, and COX-2 enzymes. The equimolar mixture showed the strongest inhibition of 5-LOX (IC₅₀ = 8.46 ± 0.92 µg/mL) and substantial inhibition of COX-1 (IC₅₀ = 15.23 ± 2.34 µg/mL) and COX-2 (IC50 = 14.53 ± 2.42 µg/mL), indicating a synergistic effect. Anticancer activity was tested on MCF-7, MDA-MB-231, and MDA-MB-436 breast cancer cell lines using the MTT assay. The thymol/carvacrol mixture demonstrated superior cytotoxicity (IC₅₀ = 0.92–1.70 µg/mL) and increased selectivity compared to cisplatin, with high selectivity indices (144.88–267.71). These results underscore the promising therapeutic potential of the thymol/carvacrol combination, particularly for its synergistic antioxidant, anti-inflammatory, and anticancer properties against breast cancer. This study paves the way for developing natural therapies against breast cancer and other conditions associated with oxidative stress and inflammation, leveraging the synergistic effects of natural compounds like thymol and carvacrol. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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10 pages, 2403 KiB

Open AccessArticle

Rosmarinic Acid Potentiates Cytotoxicity of Cisplatin against Colorectal Cancer Cells by Enhancing Apoptotic and Ferroptosis

by Jhen-Yu Huang, Ta-Wen Hsu, Yu-Ru Chen and Shao-Hsuan Kao

Life 2024, 14(8), 1017; https://doi.org/10.3390/life14081017 - 15 Aug 2024

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Abstract

Rosmarinic acid (RA) has demonstrated anticancer effects on several types of malignancies. However, whether RA promotes the anticancer effect of cisplatin on colorectal cancer cells remains sketchy. This study aimed to explore whether RA potentiates the cytotoxicity of cisplatin against colon cancer cells [...] Read more.

Rosmarinic acid (RA) has demonstrated anticancer effects on several types of malignancies. However, whether RA promotes the anticancer effect of cisplatin on colorectal cancer cells remains sketchy. This study aimed to explore whether RA potentiates the cytotoxicity of cisplatin against colon cancer cells and the underlying mechanism. Cell viability, cell cycle progression, and apoptosis was evaluated using sulforhodamine B (SRB) assay, flow cytometric analysis, and propidium iodide/Annexin V staining, respectively. Western blotting was utilized to analyze signaling pathways. Our findings showed that RA significantly enhanced the inhibitory effect on cell viability and the induction of apoptosis on the colon cancer cell lines DLD-1 and LoVo. Signaling cascade analysis revealed that the combination of RA and cisplatin jointly induced Bax and caspase activation while downregulating Bcl-2, glutathione peroxidase 4 (GPX4), and SLC7A11 in DLD-1 cells. Moreover, caspase inhibitor and ferroptosis inhibitor significantly reversed the inhibition of cell viability in response to RA combined with cisplatin. Collectively, these findings demonstrate that RA enhances the cytotoxicity of cisplatin against colon cancer cells, attributing to the promotion of apoptosis and ferroptosis. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Inhibitory effects of RA and the combination of RA with cisplatin on cell viability of colon cancer cells. Cells were treated with RA, cisplatin (Cis), or their combinations for 24 h, followed by subjecting the treated cells to SRB assay to evaluate cell viability. Cell viability was presented as percentage of control. (A) Effect of RA on cell viability of DLD-1 ad LoVo cells, (B) Effect of cisplatin on cell viability of DLD-1 ad LoVo cells, and (C,D) Effect of RA combined with cisplatin on cell viability of DLD-1 and LoVo cells. For (A,B) *, **, and ***, p < 0.05, p < 0.01, and p < 0.005 vs. control. For (C,D), # and ##, p < 0.05 and p < 0.01 vs. control, respectively. * and **, p < 0.05 and p < 0.01 vs. 20 μM cisplatin treatment alone, respectively. Full article ">Figure 2
Effects of RA, cisplatin, and the combination of cisplatin with RA on cell cycle distribution in colon cancer cells. Cells were treated with (A) 400 μM RA alone or (B) cisplatin (Cis), RA, or their combinations for 24 h, and then the treated cells were fixed and stained with PI. Flow cytometric analysis was used to determine the ratios of cells in individual cell cycle phase. a and b, p < 0.05 vs. control (-) and 20 μM cisplatin treatment alone, respectively. Full article ">Figure 3
Induction of apoptosis by RA, cisplatin, and the combination of cisplatin with RA in colon cancer cells. Cells were treated with (A) 400 μM RA alone or (B) cisplatin (Cis), RA, or their combinations for 24 h. The cells that received treatment were fixed, stained with PI and Annexin V-FITC (ANXV), and then analyzed using flow cytometry to calculate the proportions of cells showing PI-positive, ANXV-positive, or both characteristics. a and b, p < 0.05 vs. control and 20 μM cisplatin treatment alone, respectively. Full article ">Figure 4
RA and its combination with cisplatin reduced antiapoptotic Bcl-2 and enhanced proapoptotic signals in DLD-1 cells. Cells were treated with RA, cisplatin, and the combination of cisplatin with RA for 24 h, and then the cells were subjected to (A,B) assess the levels of antiapoptotic Bcl-2 and proapoptotic Bax, cleaved (clv.) caspase-9, and clv. caspase-3 by Western blotting. Relative quantitation was conducted by using densitometric analysis, and the results are shown in (C–F). Level of β-actin was used as internal control. #, p < 0.05 vs. control. ***, p < 0.005 vs. 20 μM cisplatin treatment alone. Full article ">Figure 5
RA and its combination with cisplatin promoted ferroptosis signals in DLD-1 cells. Cells were treated with RA, cisplatin, or the combination of cisplatin with RA for 24 h, and then the cells were subjected to (A,B) assess the levels of ferroptosis-related components by Western blotting. Relative quantitation was conducted by using densitometric analysis, and the results are shown in (C–E). Level of β-actin was used as internal control. #, p < 0.05 vs. control. * and ***, p < 0.05 and p < 0.005 vs. 20 μM cisplatin treatment alone, respectively. Full article ">Figure 6
Involvement of apoptosis and ferroptosis in the cytotoxicity of cisplatin combined with RA against colon cancer cells. (A) DLD-1 and (B) LoVo cells were pretreated with Z-VAD-FMK (Z-VAD) or ferrostatin-1 (Fer-1), treated with cisplatin (Cis) or cisplatin combined with RA for 24 h, and then subjected to cell viability assessment by SRB assay. Cell viability was presented as percentage of control. *, **, and ***, p < 0.05, p < 0.01, and p < 0.005, respectively. Full article ">

17 pages, 4559 KiB

Open AccessArticle

Wound Healing Potential of a Novel Sedum Species: S. album Murales

by Francesca Truzzi, Elettra Frassineti, Camilla Tibaldi, Eros D’Amen and Giovanni Dinelli

Life 2024, 14(8), 958; https://doi.org/10.3390/life14080958 - 30 Jul 2024

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Abstract

Natural wound healing products are in increased demand. The potential for unexplored Sedum species in wound healing was discovered based on benefits of the genus reported in traditional medicine. The objectives were to screen ten Sedum species for wound healing, to ascertain the [...] Read more.

Natural wound healing products are in increased demand. The potential for unexplored Sedum species in wound healing was discovered based on benefits of the genus reported in traditional medicine. The objectives were to screen ten Sedum species for wound healing, to ascertain the optimal harvest period using the five best, and finally to investigate effects of extraction protocols on wound healing using the most promising species. Different protocols were used to extract leaf polyphenol and mucilage content. Wound healing was assessed from L929 fibroblast migration. April was the optimal harvest month for wound healing efficacy, whereas the highest polyphenol content and antioxidant activity were evident in September and November. S. album Murales (ALBU), the best candidate, was then compared with S. telephium (TELE), which is well recognized in skin care. The mucilage-containing aqueous extract of ALBU was shown for the first time to induce the highest fibroblast migration after 24 h, not evident in TELE. Moreover, functional constituents contained within the absolute acetone- and isopropanol-containing polyphenol pools from ALBU induced significantly higher migration compared to TELE. A prototype cream, containing the water- and solvent-extracted bioactive compounds was effective at inducing fibroblast migration at 24 h in ALBU. The potential of ALBU in wound healing was evidenced and warrants further investigation. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Graphical abstract

Graphical abstract
Full article ">Figure 1
Percentage wound closure by L929 fibroblast migration following 4 h exposure to 10 µg/mL gallic acid equivalents (GAE) from 10 Sedum species, respectively, compared to the untreated control (CTRL). Statistical analysis was performed with the letters (a, b, c, d) representing significant differences between treatments as determined by one-way ANOVA and the Turkey-Kramer test at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. acre (ACRE), S. acre yellow (ACREY), S. album Murales (ALBU), S. herbstfreude (HERB), S. hispanicum (HISP), S. montanum (MONT), S. reflexum (REFL), S. sediforme, (SEDI) S. spectabile (SPECT) and S. telephium (TELE). Full article ">Figure 2
(A) Percentage wound closure by fibroblast migration following a 4 h exposure to 10 µg/mL GAE, representing the average of harvests conducted over five months for five Sedum species, compared to the CTRL and (B) percentage wound closure by fibroblast migration following a 4 h exposure to 10 µg/mL GAE, representing the average of five Sedum species, for each harvest month. (C) Fibroblast viability (MTT assay), expressed as percentage of the untreated CTRL (100%) for five Sedum species and five harvest months. (D) Polyphenol content (expressed in mg GAE/g) in relation to harvest month for each of the five Sedum species and (E) FRAP activity (expressed in μmol Fe2+/g) in relation to harvest month for each of the five Sedum species. (A,B,D,E) Statistical analysis was performed with the letters (a, b, c, d … o) representing significant differences between treatments as determined by one-way ANOVA and the Turkey-Kramer test at the 95% confidence level (p < 0.05). (C) Statistical analysis was performed using a two-way ANOVA at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. acre (ACRE), S. acre yellow (ACREY), S. album Murales (ALBU), S. herbstfreude (HERB) and S. telephium (TELE). Full article ">Figure 3
(A) Polyphenol content (expressed in mg GAE/g), (B) DPPH antiradical activity (expressed in %RSC), (C) FRAP activity (expressed in μmol Fe2+/g) and (D) mucilage content in TELE and ALBU (expressed in %gr). Extracts were performed in water (W), acetone (A), isopropanol (I), with mixed (MIX) extracts containing the three extraction volumes in an equivalent ratio (1:1:1). Statistical analysis was performed with the letters (a, b, c, d, e) representing significant differences between treatments as determined by one-way ANOVA and the Turkey-Kramer test at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. album Murales (ALBU), and S. telephium (TELE). Full article ">Figure 4
(A) TELE- and (B) ALBU-induced fibroblast vitality (Blue Trypan assay) compared to the untreated control (CTRL). Extracts were performed in water (W), acetone (A), isopropanol (I), with mixed (MIX) extracts containing the three extraction volumes in an equivalent ratio (1:1:1). All extracts were prepared at different extract: DMEM dilutions (1:30, 1:40, 1:50, 1:100), and the cream at a 1:100 dilution. Extracts were exposed to the cells for 24 h. Statistical analysis was performed with the number of stars **** representing significant differences between treatments as determined by one-way ANOVA at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. album Murales (ALBU), and S. telephium (TELE). Full article ">Figure 5
(A,B) Fibroblast migration percentages induced by mixed (MIX) extracts from ALBU and TELE after 4, 24 and 48 h compared to the control (CTRL). (C,E) Microscope images (×20 magnification) portraying fibroblast migration induced by extracts of TELE and ALBU, extracted in water (W), acetone (A), isopropanol (I), with MIX extracts containing the three extraction volumes in an equivalent ratio (1:1:1) after 24 h. (D,F) Quantification of fibroblast migration derived from the images in (C,E). All experiments were prepared at an extract: DMEM dilution of 1:100. Statistical analysis was performed with the number of stars, *, **, ***, **** representing significant differences between treatments as determined by one-way ANOVA at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. album Murales (ALBU), and S. telephium (TELE). Full article ">Figure 6
(A) Statistical quantification of fibroblast migration percentages induced by mixed (MIX) extracts from ALBU and TELE after 24 h compared to the control (CTRL). (B) Microscope images (x 20 magnification) portraying fibroblast migration induced by the MIX, containing constituents extracted in water (W), acetone (A), isopropanol (I) in a 1:1:1 ratio and then exposed to the cells at an extract: DMEM dilution of 1:100. Statistical analysis was performed with the number of stars, ** representing significant differences between treatments as determined by one-way ANOVA at the 95% confidence level (p < 0.05). Abbreviations for Sedum species: S. album Murales (ALBU), and S. telephium (TELE). Full article ">

15 pages, 2344 KiB

Open AccessArticle

Piceatannol Upregulates SIRT1 Expression in Skeletal Muscle Cells and in Human Whole Blood: In Vitro Assay and a Randomized, Double-Blind, Placebo-Controlled, Parallel-Group Comparison Trial

by Kenta Tanaka, Shinpei Kawakami, Sadao Mori, Takumi Yamaguchi, Eriko Saito, Yuko Setoguchi, Yuko Matsui, Eisaku Nishimura, Shukuko Ebihara and Toshihiro Kawama

Life 2024, 14(5), 589; https://doi.org/10.3390/life14050589 - 5 May 2024

Viewed by 2392

Abstract

Piceatannol (PIC), a polyphenol abundant in passion fruit seeds, is reported to promote fat metabolism. This study investigated whether PIC affects sirtuin 1 (SIRT1) expression and metabolic factors in C2C12 skeletal muscle cells. C2C12 myotubes were stimulated with PIC, and alterations in gene [...] Read more.

Piceatannol (PIC), a polyphenol abundant in passion fruit seeds, is reported to promote fat metabolism. This study investigated whether PIC affects sirtuin 1 (SIRT1) expression and metabolic factors in C2C12 skeletal muscle cells. C2C12 myotubes were stimulated with PIC, and alterations in gene expression, protein levels, mitochondrial DNA content, and fatty acid levels were assessed using real-time PCR, Western blotting, and Nile red staining. Furthermore, we examined changes in SIRT1 expression following the consumption of a test food containing 100 mg PIC for 2 weeks among adults with varying age and body mass index ranges. Both PIC and passion fruit seed extract induced SIRT1 expression in C2C12 myotubes to a greater extent than resveratrol. PIC also increased the expression of genes associated with mitochondrial biogenesis and fatty acid utilization, increased mitochondrial DNA content, and suppressed oleic acid-induced fat accumulation. Moreover, participants who consumed PIC exhibited significantly higher SIRT1 mRNA expression in whole blood compared to those in the placebo group. These findings suggest that PIC induces SIRT1 expression both in vitro and in the human body, which may promote mitochondrial biosynthesis and fat metabolism. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Figure 1
Effect of piceatannol (PIC) on SIRT1 expression in C2C12 myotubes. (A) Cells were exposed to 0.1% DMSO (CON; white bar) or PIC at concentrations of 10, 20, and 50 µM (black bar) for 6 h (n = 6), and Sirt1 mRNA expression was measured and normalized to that of Gapdh. *** p < 0.001 vs. CON (Dunnett’s test). (B) Cells were exposed to 0.1% DMSO (CON; white bar) or passion fruit seed extract (PFSE, containing 20 μM PIC; black bar) for 6 h (n = 8), and Sirt1 mRNA expression was measured and normalized to that of Gapdh. *** p < 0.001 vs. CON (Student’s t-test). (C) Cells were exposed to 0.1% DMSO (CON; white bar), 50 µM PIC (black bar), or 50 µM RES (gray bar) for 6 h (n = 4), and Sirt1 mRNA expression was measured and normalized to that of Gapdh. Significant differences were identified using Tukey’s HSD test. Different letters indicate significance (p < 0.05). (D) Following a 24 h treatment with 0.1% DMSO (CON; white bar), 50 µM PIC (black bar), or 50 µM RES (gray bar), the relative SIRT1 protein level was normalized to that of GAPDH (n = 6). The right panel of (D) depicts a typical blot image showing treatments with the control, 50 µM PIC, and 50 µM RES. Tukey’s HSD test was utilized to ascertain significant differences between treatments, with different letters indicating significance (p < 0.05). Full article ">Figure 2
Effect of piceatannol (PIC) on mitochondria-related gene expression and mtDNA copy number in C2C12 myotubes. (A) Cells were exposed to 0.1% DMSO (CON; white bar) or 50 µM PIC (black bar) for 24 h (n = 5–6). Mitochondrial gene mRNA levels were quantified and normalized to those of Gapdh. (B) Cells were exposed to 0.1% DMSO (CON; white bar) or 20 µM PIC (black bar) for 48 h (n = 7). The mitochondrial-to-nuclear DNA ratio (mtDNA/nDNA) was determined via qPCR after total DNA extraction. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. CON (Student’s t-test). Full article ">Figure 3
Effect of PIC on Ho-1 and Nqo-1 mRNA expression in C2C12 myotubes. Cells were exposed to 0.1% DMSO (CON; white bar), 50 µM PIC (black bar), or 50 µM RES (gray bar) for 6 h (n = 4), and Ho-1 and Nqo-1 mRNA levels were quantified and normalized to those of Gapdh. Significant differences were identified using Tukey’s HSD test, with different letters indicating significance (p < 0.05). Full article ">Figure 4
Effect of piceatannol (PIC) on the mRNA level of fatty acid utilization genes and fatty acid accumulation in C2C12 myotubes. (A) Cells were exposed to 0.1% DMSO (white bar) or 50 µM PIC (black bar) for 24 h (n = 5–6). Gene expression related to fatty acid utilization was quantified and normalized to that of Gapdh. * p < 0.05. (B) Cells were treated with 1 mM oleic acid (OA) or left untreated for 1 h. Subsequently, cells were incubated with or without 50 μM PIC for 48 h. Cells in the control group (CON) were treated with 0.1% DMSO, without OA treatment. Cells in the OA group were treated with 1 mM OA, whereas those in the OA+PIC group were treated with both 1 mM OA and 50 μM PIC. Intracellular lipid droplets were quantified using Nile red staining. Significant differences were identified using Tukey’s HSD test, with different letters indicating significance (p < 0.05). Full article ">Figure 5
Flow chart of the clinical trial. Full article ">

15 pages, 1364 KiB

Open AccessEditor’s ChoiceArticle

Deciphering the Systemic Impact of Herbal Medicines on Allergic Rhinitis: A Network Pharmacological Approach

by Sa-Yoon Park, Yoon Yeol Lee, Min Hee Kim and Chang-Eop Kim

Life 2024, 14(5), 553; https://doi.org/10.3390/life14050553 - 25 Apr 2024

Cited by 4 | Viewed by 1571

Abstract

Allergic rhinitis (AR) is a systemic allergic disease that has a considerable impact on patients’ quality of life. Current treatments include antihistamines and nasal steroids; however, their long-term use often causes undesirable side effects. In this context, traditional Asian medicine (TAM), with its [...] Read more.

Allergic rhinitis (AR) is a systemic allergic disease that has a considerable impact on patients’ quality of life. Current treatments include antihistamines and nasal steroids; however, their long-term use often causes undesirable side effects. In this context, traditional Asian medicine (TAM), with its multi-compound, multi-target herbal medicines (medicinal plants), offers a promising alternative. However, the complexity of these multi-compound traits poses challenges in understanding the overall mechanisms and efficacy of herbal medicines. Here, we demonstrate the efficacy and underlying mechanisms of these multi-compound herbal medicines specifically used for AR at a systemic level. We utilized a modified term frequency–inverse document frequency method to select AR-specific herbs and constructed an herb–compound–target network using reliable databases and computational methods, such as the Quantitative Estimate of Drug-likeness for compound filtering, STITCH database for compound-target interaction prediction (with a high confidence score threshold of 0.7), and DisGeNET and CTD databases for disease-gene association analysis. Through this network, we conducted AR-related targets and pathway analyses, as well as clustering analysis based on target-level information of the herbs. Gene ontology enrichment analysis was conducted using a protein–protein interaction network. Our research identified 14 AR-specific herbs and analyzed whether AR-specific herbs are highly related to previously known AR-related genes and pathways. AR-specific herbs were found to target several genes related to inflammation and AR pathogenesis, such as PTGS2, HRH1, and TBXA2R. Pathway analysis revealed that AR-specific herbs were associated with multiple AR-related pathways, including cytokine signaling, immune response, and allergic inflammation. Additionally, clustering analysis based on target similarity identified three distinct subgroups of AR-specific herbs, corroborated by a protein–protein interaction network. Group 1 herbs were associated with the regulation of inflammatory responses to antigenic stimuli, while Group 2 herbs were related to the detection of chemical stimuli involved in the sensory perception of bitter taste. Group 3 herbs were distinctly associated with antigen processing and presentation and NIK/NF-kappa B signaling. This study decodes the principles of TAM herbal configurations for AR using a network pharmacological approach, providing a holistic understanding of drug effects beyond specific pathways. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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Figure 1
Representative herb–compound–target network for AR-related genes. Full article ">Figure 2
Herb–target network for AR-specific herbs. The green nodes represent the herbs and ends of the arrowheads for the predicted targets. As a result of filtering the compounds by the quantitative estimate of drug-likeness (QED) standard (QED > 0.35), platycodon grandiflorum was excluded because no active compound was included. Full article ">Figure 3
Herb clusters of the AR-specific herbs. (A) Cosine similarity of AR-specific herbs using compound information. (B) Cosine similarity of AR-specific herbs using target information. Three blue boxes indicate herb groups obtained from the hierarchical clustering analysis. Full article ">Figure 4
Protein–protein interaction (PPI) network analysis of AR-related herb clusters. (A) The PPI network was built with the targets of the AR-specific herbs. After constructing the PPI network, the modules were classified using the Louvain method. The percentage occupied by each color-coded module is shown in the legend. (B) The PPI network of group 1 herbs (ephedra intermedia, panax ginseng, asiasarum sieboldii, and bupleurum falcatum) is shown in red. (C) The PPI network of group 2 herbs (cinnamomum cassia, angelica decursiva, Aralia continentalis, pueraria lobata, poria cocos, glycyrrhiza glabra, astragalus membranaceus, saposhnikovia divaricata) is shown in dark blue. (D) The PPI network of group 3 herb (pinellia ternata) is shown in blue. AR = allergic rhinitis. Full article ">

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19 pages, 2073 KiB

Open AccessReview

A Natural Approach to the Prevention and Treatment of Gingivitis and Periodontitis: A Review of Pomegranate’s Bioactive Properties

by Georgiana Ioana Potra Cicalău, Laura Grațiela Vicaș, Gabriela Ciavoi, Timea Claudia Ghitea, Nagy Csaba, Roxana Alexandra Cristea, Florina Miere (Groza) and Mariana Ganea

Life 2024, 14(10), 1298; https://doi.org/10.3390/life14101298 - 13 Oct 2024

Viewed by 1546

Abstract

(1) Background: This systematic review explores the bioactive properties of Punica granatum (pomegranate) and its potential applications in the prevention and treatment of gingivitis, periodontitis, and other oral diseases. (2) Methods: A comprehensive literature search was conducted using PubMed and Google Scholar, focusing [...] Read more.

(1) Background: This systematic review explores the bioactive properties of Punica granatum (pomegranate) and its potential applications in the prevention and treatment of gingivitis, periodontitis, and other oral diseases. (2) Methods: A comprehensive literature search was conducted using PubMed and Google Scholar, focusing on pomegranate and oral diseases. Inclusion criteria included studies evaluating the effects of pomegranate on oral health, while exclusion criteria eliminated non-peer-reviewed and non-English articles. This review aims to assess the efficacy of pomegranate extracts as a natural alternative to synthetic pharmaceuticals in oral health care. A structured search strategy included key terms such as “pomegranate”, “oral health”, “gingivitis”, and “periodontitis”. A total of 125 relevant references were reviewed to identify the most pertinent findings. (3) Results: The results indicate that pomegranate extracts have demonstrated efficacy in reducing plaque, inhibiting harmful oral microorganisms, and promoting overall oral health. Furthermore, clinical studies highlight the potential of pomegranate-based products, such as mouthwashes and gels, as viable alternatives to conventional pharmaceuticals, particularly in resource-limited settings. However, the review also notes the need for further research, particularly in the form of clinical trials, to establish optimal formulations and long-term safety. (4) Conclusions: Pomegranate presents a promising, natural solution for preventing and treating gingivitis and periodontitis. Further studies should focus on long-term effects and clinical efficacy. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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18 pages, 1371 KiB

Open AccessReview

Therapeutic Potential of Pomegranate Extract for Women’s Reproductive Health and Breast Cancer

by Jung Yoon Jang, Donghwan Kim, Eunok Im and Nam Deuk Kim

Life 2024, 14(10), 1264; https://doi.org/10.3390/life14101264 - 3 Oct 2024

Viewed by 2966

Abstract

Pomegranate extract has potential benefits for women’s reproductive health, including fertility enhancement, menstrual cycle regulation, pregnancy support, and polycystic ovary syndrome (PCOS) treatment. It possesses antioxidant properties, reducing oxidative stress and improving fertility. Pomegranate extract may help regulate hormonal imbalances and promote regular [...] Read more.

Pomegranate extract has potential benefits for women’s reproductive health, including fertility enhancement, menstrual cycle regulation, pregnancy support, and polycystic ovary syndrome (PCOS) treatment. It possesses antioxidant properties, reducing oxidative stress and improving fertility. Pomegranate extract may help regulate hormonal imbalances and promote regular menstrual cycles. The extract’s rich nutrient profile supports placental development and fetal growth and may reduce the risk of preterm birth. Additionally, pomegranate extract shows promise in improving insulin sensitivity and reducing inflammation and oxidative damage in PCOS. Some studies suggest its potential anticancer properties, particularly against breast cancer. However, further research, including human clinical trials, is necessary to establish its effectiveness and safety. The current evidence is limited and primarily based on in vitro studies, animal studies, and clinical trials. This review provides a comprehensive summary of the benefits of pomegranate extract for women’s reproductive health and breast cancer, serving as a reference for future research. Full article

(This article belongs to the Special Issue Advances in the Biomedical Applications of Plants and Plant Extracts)

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