芍药甘草汤通过JAK2/STAT3信号通路调控小胶质细胞极化的机制

Mechanism of Shaoyao Gancao Decoction(芍药甘草汤)in the Modulation of Microglial Polarization through JAK2/STAT3 Signaling Pathway

目的研究芍药甘草汤含药血清对BV-2细胞极化的调控作用与机制。方法采用LPS激活BV-2细胞制备小胶质细胞炎症模型,细胞随机分为空白组、模型组(1 mg/L LPS)、芍药甘草汤含药血清低、中、高剂量组(5%、10%、25%)。CCK-8法检测细胞活力;酶联免疫吸附测定法(ELISA)检细胞中IL-1β、IL-6、TNF-α、iNOS的表达;Real-time PCR检测Arg-1、CD68、SOCS3 mRNA表达;蛋白免疫印迹法(Western blot)检测JAK2、p-JAK2、STAT3、p-STAT3蛋白表达水平。结果与空白组比较,模型组BV-2细胞活力和Arg-1 mRNA表达显著降低(P<0.01),IL-1β、IL-6、TNF-α、iNOS和CD68、SOCS3 mRNA表达以及JAK2、p-JAK2、STAT3、p-STAT3蛋白表达明显升高(P<0.01)。与模型组比较,芍药甘草汤含药血清能提高BV-2细胞活力和Arg-1 mRNA表达(P<0.01),而IL-1β、IL-6、TNF-α、iNOS含量,CD68、SOCS3 mRNA表达以及JAK2、p-JAK2、STAT3、p-STAT3蛋白表达明显降低(P<0.05)。结论芍药甘草汤含药血清可促进小胶质细胞由M1型极化为M2型,改善神经炎症反应,其抗炎作用可能与下调JAK2/STAT3信号通路有关。

Objective To investigate the function and regulatory mechanism of Shaoyao Gancao Decoction(芍药甘草汤,SGD)-containing serum on the polarization of BV-2 cells.Methods Microglia inflammation model was established by inducing BV-2 cells with lipopolysaccharide(LPS).All cells were divided into the blank,model(1 mg/L LPS),and SGD-contained serum low,medium,and high-dose groups(5%,10%and 25%).Cell viability was evaluated utilizing the CCK-8 assay.The expressions of IL-1β,IL-6,TNF-αand iNOS were detected via ELISA.Real-time PCR was used to detect Arg-1,CD68,and SOCS3 mRNA expression.The proteins JAK2,p-JAK2,STAT3 and p-STAT3 were investigated by Western blot.Results Compared with the blank group,BV-2 cell viability significantly decreased,Arg-1 mRNA expression level were significantly reduced(P<0.01),the expression of IL-1β,IL-6,TNF-α,iNOS were markedly increased,CD68,SOCS3 mRNA expression and JAK2,p-JAK2,STAT3,p-STAT3 protein expression were increased in the model group(P<0.01).Compared with the model group,SGD containing serum increased the BV-2 cell viability and Arg-1 mRNA expression(P<0.01),decreased IL-1β,IL-6,TNF-α,iNOS expression,reduced CD68,SOCS3 mRNA and JAK2,p-JAK2,STAT3,p-STAT3 protein levels(P<0.05).Conclusion SGD-containing serum promotes the polarization of M1 microglia to an M2 phenotype,improves neuroinflammation,and the mechanism may be related to the downregulation of JAK2/STAT3 signaling pathway.