白藜芦醇对脂多糖诱导的人肺上皮细胞焦亡的保护机制研究

Study on Protective Mechanism of Resveratrol against Pyroptosis induced by Lipopolysaccharide in Human Lung Epithelial Cells

目的:探究白藜芦醇对脂多糖(LPS)诱导的人肺上皮细胞(又称BEAS-2B)增殖、炎症因子释放和焦亡的影响。方法:用CCK-8法检测LPS和LPS与白藜芦醇联用对BEAS-2B细胞增殖的影响;采用ELISA法检测细胞上清炎症因子肿瘤坏死因子(TNF-α)、白介素-6(IL-6)、白介素-1β(IL-1β)和白介素-18(IL-18)表达以及qPCR和Western blot检测焦亡基因NLRP3、Gasdermin D、Caspase-1、ELAVL1的表达;分析其对细胞活力、BEAS-2B细胞中细胞因子的含量及其焦亡相关基因、相关蛋白表达的影响。结果:LPS可以抑制BEAS-2B细胞的增殖,同时可以促进炎症因子的表达,经qPCR和Western blot检测结果表明LPS可以促进NLRP3、Gasdermin D、Caspase-1、ELAVL1基因的表达;其经用白藜芦醇处理48 h后,可以有效逆转LPS所引起的细胞增殖受抑制和炎症因子高表达的现象;此外,NLRP3、Gasdermin D、Caspase-1、ELAVL1的表达也部分受抑制。结论:白藜芦醇通过减少炎症因子的释放和NLRP3、Gasdermin D、Caspase-1、ELAVL1表达对LPS诱导的BEAS-2B焦亡起到一定的保护作用。

Objective:To explore the influence of resveratrol on the proliferation,release of inflammatory cytokines and pyroptosis of human lung epithelial cells(also known as BEAS-2B)induced by LPS.Methods:The proliferation of BEAS-2B cells treated with lipopolysaccharide(LPS)or LPS combined with resveratrol was assayed by CCK-8 method;expression of inflammatory cytokines TNF-α,IL-6,IL-1βand IL-18 in cell supernatant were measured with ELISA,qPCR and Western blot were used to detect the expressions of genes related with pyroptosis NLRP3,Gasdermin D,Caspase-1,and ELAVL1;their influence on cell viability,cytokine content,expression of pyroptosis related genes and proteins in BEAS-2B cells were analyzed.Results:LPS could inhibit the proliferation of BEAS-2B cells;at the same time,the expression of Inflammatory cytokines can be promoted.The results of qPCR and Western blot showed that LPS could promote the expression of NLRP3,Gasdermin D,Caspase-1,ELAVL1.It was treated with resveratrol for 48 hours the phenomenon of inhibited cell proliferation and high expression of inflammatory factors caused by LPS can be effectively reversed.In addition,the expressions of NLRP3,Gasdermin D,Caspase-1,and ELAVL1 were also partially suppressed.Conclusion:Resveratrol can protect LPS-induced pyroptosis in BEAS-2B cells by reducing the release of inflammatory cytokines and the expression of NLRP3,Gasdermin D,Caspase-1,and ELAVL1.