Abstract
Background
Oxidative stress is one of the most important factors affecting large-scale breeding, especially the performance of pigs. Oxidative stress plays a role by affecting various genes in pigs, which can cause serious body damage, functional degradation and reduce production performance.
Objective
The purpose of this study was to investigate the effect of Toll like receptor 9 (TLR9) pathway on IPEC-J2 cells under oxidative stress and to provide reference for the growth development of Dapulian pigs.
Methods
In this study, Diquat was used as a source of oxidative stress to study the effects on Dapulian pigs by detecting relevant indicators. Then the IPEC-J2 cells were selected to verify the TLR9 signaling pathway in oxidative stress.
Results
Compared with the control group, superoxide dismutase (SOD) in experimental group decreased significantly, malondialdehyde (MDA) was significantly increased, accompanied by inflammatory reaction, and inflammatory factors were significantly increased in the experimental group. Oxidative stress model was constructed by H2O2 incubating IPEC-J2 cells. The interference and overexpression vectors of TLR9 and myeloid differentiation primary response protein 88 (MyD88) were constructed to detect the activity of antioxidant enzymes and related proteins. The results showed that overexpression of TLR9 enhanced the activity of antioxidant enzymes, decreased the secretion of inflammatory factors, and decreased the activity of MDA，reactive oxygen species (ROS); the results were opposite after TLR9 interference. This study also showed that H2O2 can activate the nuclear factor-κB (NF-κB) pathway and promote the translocation of NF-κB into the nucleus. After co-transfection with TLR9 and MyD88, the results showed that TLR9 regulated the expression of NF-κB through MyD88.
Conclusion
The study showed that TLR9 pathway had a significant positive effect on antioxidant.
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Data availability
The data are available from the corresponding author upon request.
References
Abraham KADS (2004) Involvement of reactive oxygen species in toll-like receptor 4-dependent activation of NF-κB. J Immunol. https://doi.org/10.1080/00397919508011369
Akyol O, Erdemli HK (2015) A new remedial approach to oxidant/antioxidant imbalance-based diseases: wet-cupping therapy. Complement Ther Med 23:633. https://doi.org/10.1016/j.ctim.2015.06.008
Cai X, Zhu L, Chen X, Sheng Y, Guo Q, Bao J et al (2016) X/XO or H2O2 induced IPEC-J2 cell as a new in vitro model for studying apoptosis in post-weaning piglets. Cytotechnology 68:713–724. https://doi.org/10.1007/s10616-014-9823-z
Castello L, Froio T, Maina M, Cavallini G, Biasi F, Leonarduzzi G et al (2010) Alternate-day fasting protects the rat heart against age-induced inflammation and fibrosis by inhibiting oxidative damage and NF-kB activation. Free Radic Bio Med 48:47–54. https://doi.org/10.1016/j.freeradbiomed.2009.10.003
Chen Z, Yuan Q, Xu G, Chen H, Lei H, Su J (2018) Effects of quercetin on proliferation and H(2)O(2)-induced apoptosis of intestinal porcine enterocyte cells. Molecules. https://doi.org/10.3390/molecules23082012
Chen Y, Chen Y, Zhang H, Wang T (2020) Pterostilbene as a protective antioxidant attenuates diquat-induced liver injury and oxidative stress in 21-day-old broiler chickens. Poultry Sci 99:3158–3167. https://doi.org/10.1016/j.psj.2020.01.021
Fentoglu O, Kirzioglu FY, Bulut MT, Kumbul DD, Kulac E, Onder C et al (2015) Evaluation of lipid peroxidation and oxidative DNA damage in patients with periodontitis and hyperlipidemia. J Periodontol 86:682–688. https://doi.org/10.1902/jop.2015.140561
Fussell KC, Udasin RG, Gray JP, Mishin V, Smith PJS, Heck DE et al (2011) Redox cycling and increased oxygen utilization contribute to diquat-induced oxidative stress and cytotoxicity in Chinese hamster ovary cells overexpressing NADPH-cytochrome P450 reductase. Free Radic Bio Med 50:874–882. https://doi.org/10.1016/j.freeradbiomed.2010.12.035
Gallelli L, Falcone D, Scaramuzzino M, Pelaia G, D’Agostino B, Mesuraca M et al (2014) Effects of simvastatin on cell viability and proinflammatory pathways in lung adenocarcinoma cells exposed to hydrogen peroxide. BMC Pharmacol Toxico. https://doi.org/10.1186/2050-6511-15-67
Helenius M, Hänninen M, Lehtinen SK, Salminen A (1996) Aging-induced up-regulation of nuclear binding activities of oxidative stress responsive NF-kB transcription factor in mouse cardiac muscle. J Mol Cell Cardiol 28:487–498. https://doi.org/10.1006/jmcc.1996.0045
Ho MF, Ingle JN, Bongartz T, Kalari KR, Goss PE, Shepherd LE et al (2017) TCL1A single-nucleotide polymorphisms and estrogen-mediated toll-like receptor-MYD88-dependent nuclear factor-kappaB activation: single-nucleotide polymorphism- and selective estrogen receptor modulator-dependent modification of inflammation and immune response. Mol Pharmacol 92:175–184. https://doi.org/10.1124/mol.117.108340
Jia R, Gu Z, He Q, Du J, Cao L, Jeney G et al (2019) Anti-oxidative, anti-inflammatory and hepatoprotective effects of Radix Bupleuri extract against oxidative damage in tilapia (Oreochromis niloticus) via Nrf2 and TLRs signaling pathway. Fish Shellf Immun 93:395–405. https://doi.org/10.1016/j.fsi.2019.07.080
Karki K, Pande D, Negi R, Khanna S, Khanna RS, Khanna HD (2014) Expression of serum toll-like receptor 9 and oxidative damage markers in benign and malignant breast diseases. DNA Cell Biol 33:630–636. https://doi.org/10.1089/dna.2014.2405
Kawai T, Akira S (2007) Signaling to NF-kappaB by toll-like receptors. Trends Mol Med 13:460–469. https://doi.org/10.1016/j.molmed.2007.09.002
Krug A, French AR, Barchet W, Fischer JAA, Dzionek A, Pingel JT et al (2004) TLR9-dependent recognition of MCMV by IPC and DC generates coordinated cytokine responses that activate antiviral NK cell function. Immunity 21:107–119. https://doi.org/10.1016/j.immuni.2004.06.007
Lin J, Jing R, Lin F, Ge W, Dai H, Pan L (2018) High tidal volume induces mitochondria damage and releases mitochondria DNA to aggravate the ventilator-induced lung injury. Front Immunol. https://doi.org/10.3389/fimmu.2018.01477
Lu T, Piao XL, Zhang Q, Wang D, Piao XS, Kim SW (2010) Protective effects of Forsythia suspensa extract against oxidative stress induced by diquat in rats. Food Chem Toxicol 48:764–770. https://doi.org/10.1016/j.fct.2009.12.018
Lykkesfeldt J, Svendsen O (2007) Oxidants and antioxidants in disease: oxidative stress in farm animals. Vet J 173:502–511. https://doi.org/10.1016/j.tvjl.2006.06.005
Ma L, Qu Y, Wen Z, Tang Z, Guan R, Xiang X (2018) Protective effect of low molecular-weight seleno-aminopolysaccharides against H2O2-induecd oxidative stress in intestinal epithelial cells. Int J Biol Macromol Struct Funct Interact 112:745–753
Osburn WO, Wakabayashi N, Misra V, Nilles T, Biswal S, Trush MA et al (2006) Nrf2 regulates an adaptive response protecting against oxidative damage following diquat-mediated formation of superoxide anion. Arch Biochem Biophys 454:7–15. https://doi.org/10.1016/j.abb.2006.08.005
Rahman I, Biswas SK, Kode A (2006) Oxidant and antioxidant balance in the airways and airway diseases. Eur J Pharmacol 533:222–239. https://doi.org/10.1016/j.ejphar.2005.12.087
Rani V, Deep G, Singh RK, Palle K, Yadav UC (2016) Oxidative stress and metabolic disorders: pathogenesis and therapeutic strategies. Life Sci 148:183–193. https://doi.org/10.1016/j.lfs.2016.02.002
Stottmeier B, Dick TP (2016) Redox sensitivity of the MyD88 immune signaling adapter. Free Radic Bio Med 101:93–101. https://doi.org/10.1016/j.freeradbiomed.2016.10.004
Toledano MB, Leonard WJ (1991) Modulation of transcription factor NF-kappa B binding activity by oxidation-reduction in vitro. Proc Natl Acad Sci USA 88:4328–4332. https://doi.org/10.1073/pnas.88.10.4328
Wang Q, Shen J, Yan Z, Xiang X, Mu R, Zhu P et al (2020) Dietary Glycyrrhiza uralensis extracts supplementation elevated growth performance, immune responses and disease resistance against Flavobacterium columnare in yellow catfish (Pelteobagrus fulvidraco). Fish Shellfish Immun 97:153–164. https://doi.org/10.1016/j.fsi.2019.12.048
Yin J, Liu M, Ren W, Duan J, Yang G, Zhao Y et al (2015) Effects of dietary supplementation with glutamate and aspartate on diquat-induced oxidative stress in piglets. PLoS ONE 10:e122893. https://doi.org/10.1371/journal.pone.0122893
Yu B, Lv M, Zheng P, Mao XB, He J, Chen DW (2012) Responses of growth performance and tryptophan metabolism to oxidative stress induced by diquat in weaned pigs. Animal 6:928–934. https://doi.org/10.1017/S1751731111002382
Yu D, Shi M, Bao J, Yu X, Li Y, Liu W (2016) Genipin ameliorates hypertension-induced renal damage via the angiotensin II-TLR/MyD88/MAPK pathway. Fitoterapia 112:244–253. https://doi.org/10.1016/j.fitote.2016.06.010
Yuan Z, Liang Z, Yi J, Chen X, Li R, Wu Y et al (2019) Protective effect of koumine, an alkaloid from gelsemium sempervirens, on injury induced by H(2)O(2) in IPEC-J2 cells. Int J Mol Sci. https://doi.org/10.3390/ijms20030754
Zhong J, Wang Z, Xie Q, Li T, Chen K, Zhu T et al (2020) Shikonin ameliorates d-galactose-induced oxidative stress and cognitive impairment in mice via the MAPK and nuclear factor-κB signaling pathway. Int Immunopharmacol 83:106491. https://doi.org/10.1016/j.intimp.2020.106491
Acknowledgements
This study was supported financially by National Key R&D Program of China (no. 2021YFD1301200), the Agricultural Animal Breeding Project of Shandong Province (no. 2020LZGC012), Shandong Province Pig Industry Technology System Project (no. SDAIT-08-02), Shandong Provincial Natural Science Foundation (no. ZR2019MC053).
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Ma, L., Geng, J., Chen, W. et al. Effects of TLR9/NF-κB on oxidative stress and inflammation in IPEC-J2 cells. Genes Genom 44, 1149–1158 (2022). https://doi.org/10.1007/s13258-022-01271-8
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DOI: https://doi.org/10.1007/s13258-022-01271-8