白藜芦醇对脂多糖诱导的人肺泡上皮细胞A549凋亡的保护作用机制

The protective mechanism of resveratrol on LPS induced human alveolar epithelial cells A549 apoptosis

目的:探讨白藜芦醇对脂多糖(lipopolysaccharide,LPS)诱导的人肺泡上皮细胞A549增殖、凋亡的影响。方法:用MTT法检测LPS对A549细胞增殖的影响;流式细胞仪检测细胞凋亡;Western blot检测凋亡相关信号通路蛋白的表达水平;实时荧光定量PCR和免疫荧光法分别在mRNA和蛋白水平检测其表达情况。结果:LPS能够以浓度梯度和时间梯度依赖的方式显著诱导肺癌细胞A549的凋亡,同时LPS能够升高细胞内活性氧的水平。Western blot结果表明LPS能够增加细胞内p53的表达水平并降低Bcl-2的表达。用白藜芦醇处理48h后,LPS所引起的细胞内活性氧水平升高的现象显著下降。Western blot结果表明白藜芦醇能够有效逆转LPS所引起的细胞内p53表达水平升高以及Bcl-2表达下降的情况。结论:白藜芦醇通过降低细胞内活性氧、p53水平以及升高Bcl-2水平对LPS所诱导的人肺泡上皮细胞A549凋亡起到保护作用。

Objective：To investigate the effects of resveratrol on LPS induced human alveolar epithelial cells A549 apoptosis and proliferation. Methods ：To detect the effect of LPS on the proliferation of A549 cells by MTr assay. Cell apoptosis was detected by flow cytometry. Western blot was applied to explore the protein levels of the apoptosis relat- ed signal pathways. RT - qPCR and immunofluorescence were used to detect the mRNA and protein levels. Results： LPS significantly induced A549 cell apoptosis in a time - and dose - dependent manner. At the same time, LPS can increase intracellular reactive oxygen species levels. Western blot showed that LPS can increase p53 protein level and decrease Bcl -2 protein level. With resveratrol treating for 48h, elevated levels of reactive oxygen species induced by LPS was significantly decreased. Conclusion： Resveratrol can effectively reverse LPS induced intracellular p53 and Bcl -2 protein levels.