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| Eicosapentaenoic acid inhibited lipopolysaccharideinduced mesangial cell pyroptosis by regulating Nrf2 pathway |
| LI Jie, FENG Ye, LU Qingxue, LIANG Junmeng |
| （Department of Rheumatology, Minzu Hospital of Guangxi Zhuang Autonomous Region, Guangxi Nanning 530001, China） |
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Abstract Objective： To explore the regulatory effect of eicosapentaenoic acid (EPA) on lipopolysaccharide (LPS)induced HBZY1 cell pyroptosis based on the Nrf2/NLRP3/GSDMD signaling pathway. Additionally, nuclear factor E2 related factor 2 (Nrf2) agonist (4OI) and nucleotidebound oligomerized domainlike receptors (NLRP3) receptor inhibitor (MCC950) were chosen to explore the regulating mechanism of EPA on Nrf2. Methods： According to different drug treatments strategy, HBZY1 cells were divided into five groups: control group, LPS group, LPS+EPA group, LPS+EPA+4OI group, and LPS+EPA+MCC950 group, in which 10 μg/mL LPS was applied to induce the cell pyroptosis in vitro, and the concentration of EPA, 4OI, MCC950 was all 200 μmol/L, the drugs were added to the culture plate and treated for 48 h. In addition, CCK8 assay, LDH release test, and TUNEL staining were performed to detect cell damage. The release of inflammatory factors IL1β and IL18 in the cell supernatant were detected by ELISA kits, protein levels of Nrf2, NLRP3, Caspase1 and GSDMD were detected by Western Blotting. Results： Cell viability assay showed that 10 μg/mL LPS stimulation for 48 hours significantly reduced the cell viability (P<005), but 20-200 μmol/L EPA had significantly protective effects (P<005). Moreover, 200 μmol/L EPA significantly attenuated the cell damage caused by LPS (P<001), reduced the release of inflammatory factors (P<001), LDH release (P<001) and DNA fragmentation, inhibited the activation of the pyroptosis, and promoted the expression of Nrf2 protein (P<001). In addition, cotreatment with 4OI and MCC950 found that 4OI can promote the inhibitory effect of EPA on pyroptosis (P<005), reduce the expression of related proteins and the release of IL1β and IL18 (P<005), and enhance the protection effect of EPA (P<001), while MCC950 has no significant on the effect of EPA. Conclusion： EPA inhibits the expression of various molecules in the pyroptosis of HBZY1 cells induced by LPS through Nrf2, and exerts a significant cytoprotective effect.
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Received: 27 March 2023
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. [J]. Journal of Jiangsu University(Medicine Edition), 2024, 34(04): 290-. |
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2024, Vol. 34 
