基于p38MAPK/Nrf2/HO-1通路探讨清达颗粒对脂多糖诱导活化的小胶质细胞抗氧化作用研究

Anti-oxidation of Qingda Granules on Lipopolysaccharide-induced Microglia Based on p38MAPK/Nrf2/HO-1 Pathway

目的观察清达颗粒(QDG)对脂多糖(LPS)诱导活化的小胶质细胞抗氧化作用,探讨其与p38MAPK/Nrf2/HO-1抗氧化信号通路的可能联系。方法体外培育BV-2小胶质细胞,采用MTT法筛选合适的药物浓度,之后采用LPS(1μg/mL)诱导炎症模型,将其分为对照组、LPS组、LPS+QDG组,并LPS+QDG组设置31.25μg/mL、62.50μg/mL、125.00μg/mL 3个质量浓度亚组,检测各组细胞活性氧(ROS)、肿瘤坏死因子-α(TNF-α)、丙二醛(MDA)以及谷胱甘肽过氧化物酶(GSH-Px)的表达情况,Western Blot法检测磷酸化p38MAPK(p-p38)、p38MAPK(p38)、转录因子Nrf2、细胞核内Nrf2、血红素加氧酶-1(HO-1)的蛋白表达情况以及p38抑制剂干预LPS诱导的炎症细胞HO-1蛋白的表达情况。结果LPS+QDG组ROS、TNF-α、MDA的释放抑制受到一定程度上促进GSH-Px的释放,并升高p-p38、细胞核内Nrf2以及HO-1蛋白的表达。p38抑制剂干预后下调了LPS+QDG组HO-1蛋白的表达。结论清达颗粒可有效减轻LPS诱导的BV-2小胶质细胞的氧化应激损伤,这可能与激活p38MAPK/Nrf2/HO-1信号转导通路有关。

Objective To observe the antioxidant effect of Qingda granules(QDG)on lipopolysaccharide(LPS)-induced activated microglia,and explore its possible connection with p38MAPK/Nrf2/HO-1 antioxidant signaling pathway.Methods BV-2 microglial cells were cultured in vitro.The appropriate drug concentration was selected by MTT method.LPS(1μg/mL)was used to induce the inflammation models,which were divided into control group,LPS group,LPS+QDG group.The LPS+QDG group was set to 31.25μg/mL,62.50μg/mL,and 125.00μg/mL.The expressions of reactive oxygen species(ROS),tumor necrosis factor alpha(TNF-α),malondialdehyde(MDA)and glutathione peroxidase(GSH-Px)in each group were detected.Western blot method was used to detect the expression of phosphorylated p38MAPK(p-p38),p38MAPK(p38),transcription factor Nrf2,nucleus Nrf2 and heme oxygenase-1(HO-1)protein expression and p38 inhibitors interfere with the expression of HO-1 protein in LPS-induced inflammatory cells.Results The release of ROS,TNF-α,and MDA were significantly inhibited,the release of GSH-Px was promoted to a certain extent,and the expressions of phosphorylated p38MAPK,Nrf2 in the nucleus,and HO-1 protein were increased in LPS+QDG group.P38 inhibitor significantly reduced the expression of HO-1 protein in LPS+QDG group.Conclusion QDG can effectively reduce the oxidative stress injury of BV-2 microglia induced by LPS,which may be related to the activation of p38MAPK/Nrf2/HO-1 signal transduction pathway.