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目的探讨人参皂苷Rg1(Rg1)对脂多糖(LPS)诱导BV-2小胶质细胞中增殖细胞核抗原(PCNA)和细胞周期蛋白D1(cyclin D1)的调控作用及其对活化的BV-2小胶质细胞增殖的调控作用。方法体外培养BV-2小胶质细胞并分为空白对照组、LPS不同时间(1,3,6,8,12 h)处理组及Rg1不同浓度(10、20和50μmol/L)预处理组。采用Western blotting检测细胞增殖相关因子PCNA蛋白和cyclin D1蛋白的表达变化;RT-PCR检测细胞增殖相关因子PCNA和cyclin D1 mRNA的表达变化;免疫荧光染色法观察细胞内cyclin D1蛋白表达变化。采用流式细胞术检测细胞周期的变化。结果 LPS诱导BV-2小胶质细胞PCNA和cyclin D1mRNA和蛋白表达上调,并呈时间依赖性;不同浓度Rg1预处理下调LPS诱导的BV-2小胶质细胞PCNA和cyclin D1的mRNA和蛋白表达。结论Rg1通过下调LPS诱导的BV-2小胶质细胞PCNA和cyclin D1的表达水平来调控细胞周期进而抑制活化的小胶质细胞的增殖。
Objective To investigate the regulatory effect of ginsenoside Rg1 on proliferating cell nuclear antigen (PCNA) and cyclin D1 in BV-2 microglia induced by lipopolysaccharide (LPS) and its effect on activated BV-2 Regulation of glial cell proliferation. Methods BV-2 microglial cells were cultured in vitro and divided into blank control group, LPS treated groups (1,3,6,8,12 h) and Rg1 pretreatment groups (10, 20 and 50 μmol / L) . The expressions of PCNA and cyclin D1 were detected by Western blotting. The expressions of PCNA and cyclin D1 mRNA were detected by RT-PCR. The expression of cyclin D1 was detected by immunofluorescence staining. Flow cytometry was used to detect cell cycle changes. Results LPS induced up-regulation of mRNA and protein expression of PCNA and cyclin D1 in BV-2 microglia cells in a time-dependent manner. Different concentrations of Rg1 pretreatment down-regulated mRNA and protein expression of PCNA and cyclin D1 in BV-2 microglial cells induced by LPS . Conclusion Rg1 can regulate the cell cycle and down-regulate the proliferation of activated microglia by down-regulating the expression of PCNA and cyclin D1 in BV-2 microglia induced by LPS.