目的:基于Nrf2通路探讨姜黄素对卡介苗(BCG)感染巨噬细胞氧化应激的抑制作用.方法:以THP-1源性巨噬细胞建立感染模型,分为4组:对照组、BCG组、BCG+姜黄素组、BCG+姜黄素+ML385组.采用活性氧(ROS)检测试剂盒,荧光显微镜下观察各组细胞ROS荧光强度;采用比色法检测细胞还原型谷胱甘肽(GSH)含量;Western blot检测Nrf2及其下游靶基因NQO1、HO-1的蛋白表达;MTT法检测各组细胞增殖率.结果:BCG感染显著增强ROS荧光强度,降低细胞GSH含量(P<0.01),抑制Nrf2及其下游靶基因NQO1、HO-1的蛋白表达,抑制细胞增殖(P<0.01);而姜黄素可显著减弱ROS荧光强度,提高GSH水平(P<0.05),促进Nrf2、NQO1、HO-1蛋白表达及细胞增殖(P<0.01);Nrf2抑制剂ML385则逆转了姜黄素的上述作用.结论:姜黄素可以通过增加Nrf2的表达,诱导下游抗氧化分子的转录从而减轻BCG诱导的巨噬细胞氧化应激.
Objective:To investigate the inhibitory effect of curcumin on oxidative stress in BCG-infected macrophages based on the Nrf2 pathway.Methods:THP-1-derived macrophages were infected.The experiment was divided into control group,BCG group,BCG+curcumin group and BCG+curcumin+ML385 group.Cellular ROS fluorescence intensity were observed under a fluores-cence microscope;Glutathione(GSH)levels were measured by Colorimetry;Western blot was used to detect the protein expressions of Nrf2,HO-1 and NQO1;MTT was used to detect the proliferation rate of macrophages.Results:BCG infection significantly enhanced ROS fluorescence intensity,reduced cell GSH content(P<0.01),inhibited protein expressions of Nrf2,HO-1 and NQO1,at the same time inhibited cell proliferation(P<0.01);curcumin significantly weakened ROS fluorescence intensity,increased GSH level(P<0.05),promoted Nrf2,HO-1 and NQO1 protein expressions and cell proliferation(P<0.01);Nrf2 inhibitor ML385 reversed the effect of curcumin.Conclusion:Curcumin can alleviate BCG-induced oxidative stress in macrophages by increasing the expression of Nrf2 and inducing the transcription of downstream antioxidant molecules.
赵剑秋;韩晓群;邓琴;杨婧;吴快英;黄欢
宜春学院,宜春 336000
基础医学
姜黄素;Nrf2;卡介苗;巨噬细胞;氧化应激
Curcumin;Nrf2;BCG;Macrophages;Oxidative stress
《中国免疫学杂志》 2023 (012)
2523-2527 / 5
本文为江西省卫健委科技计划项目(202311177).
10.3969/j.issn.1000-484X.2023.12.011
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