淫羊藿苷对脂多糖+γ-干扰素复合诱导的BV2小胶质细胞的调节作用

Effects of icariin on BV2 microglia induced by lipopolysaccharide+interferon-γ

目的研究淫羊藿苷(icariin,ICA)对细菌脂多糖(lipopolysaccharide,LPS)加γ-干扰素(interferon-γ,IFN-γ)复合诱导小鼠小胶质细胞系BV2细胞炎性反应模型的调节作用。方法采用LPS+IFN-γ复合诱导小鼠小胶质细胞系BV2细胞,构建拟神经炎性反应的小胶质细胞活化的体外细胞模型。将传代培养的BV2细胞分为对照组、LPS+IFN-γ模型组(模型组)以及不同浓度ICA 1、2、4、8、16、32、64、128、256μmol/L干预组。采用CCK8法检测各组细胞存活率,采用ELISA法检测各组细胞培养上清中一氧化氮(NO)水平,采用免疫荧光染色技术观察BV2细胞CD16/32蛋白表达情况。结果与对照组比较,模型组BV2细胞培养上清中NO水平明显高〔(16.46±0.73)μmol/L vs.(2.55±0.41)μmol/L;t=16.59,P<0.01〕,与模型组比较,ICA 16、32、64μmol/L干预组其NO表达水平明显降低〔分别为(13.76±0.55)、(12.66±0.51)、(10.60±0.46)μmol/L;P<0.05,P<0.01,P<0.01〕。与对照组比较,模型组CD16/32表达增加〔(5823.00±109.99)vs.(118.60±89.04);t=48.77,P<0.01〕;与模型组比较,ICA 32、64μmol/L干预组小胶质细胞CD16/32蛋白表达均明显降低(分别为3327.67±126.73、2725.00±139.37,均P<0.01)。结论ICA能够显著抑制活化的小胶质细胞释放NO,抑制小胶质细胞CD16/32表达,对LPS+IFN-γ复合诱导BV2小胶质细胞具有调节作用。

Objective To investigate the effects of icariin(ICA)on cell model of neuroinflammation,which were mice BV2 microglial cells induced by lipopolysaccharide(LPS)and interferon-γ(IFN-γ).Methods Mice microglia BV2 cells were induced by LPS+IFN-γto construct an in vitro cell model of microglia activation.BV2 cells were divided into acontrol group(CTRL),an LPS+IFN-γmodel groupand intervention groups with different concentrations of ICA 1,2,4,8,16,32,64,128,256μmol/L.CCK8 method was used to detect the cell survival rate of each group.The level of nitric oxide(NO)in the cell culture supernatant of each group was detected by ELISA.CD16/32 protein expression of BV2 cells was observed by immunofluorescence staining.Results The NO level in the culture supernatant of BV2 cells in the model group was significantly higher when compared with the control group[(16.46±0.73)μmol/L vs.(2.55±0.41)μmol/L;t=16.59,P<0.01].The intervention groups with ICA 16,32 and 64μmol/L had lower levels of NO[(13.76±0.55),(12.66±0.51)and(10.60±0.46)μmol/L]than the model group(P<0.05,P<0.01 and P<0.01).When compared with the control group,the model group had more CD16/32 protein(5823.00±109.99 vs.118.6±89.04;t=48.77,P<0.01).The expressions of CD16/32 protein of microglia in the intervention groups with ICA 32 and 64μmol/L(3327.67±126.73,2725.00±139.37)were lower than that in the model group(both P<0.01).Conclusions ICA can significantly inhibit the production of NO and inhibit the expression of CD16/32 in activated microglia.ICA has a regulatory effect on BV2 cells induced by LPS+IFN-γ.