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 possible role of ginsenoside Rg1( Rg1) mediated in proliferating cell nuclear antigen( PCNA) and cyclin D1 of BV-2 cells stimulated by lipopolysaccharide( LPS) and to investigate the effect of Rg1 regulating proliferation in activated BV-2 cells. Methods BV-2 cells were divided into control group; LPS treatment groups with LPS stimulated for 1,3,6,8,12 hours,respectively and the Rg1 pretreatment groups in which the cells were pretreated with Rg1( 10,20 and 50μmol / L) prior to LPS stimulate. The protein level and mRNA expression of PCNA and cyclin D1 were detected by RT-PCR and Western blotting,respectively. The expressions of cyclin D1 were detected by immunofluorescence staining. The cell cycle was detected by flow cytometry. Results The protein level and mRNA expression of PCNA and cyclin D1 in BV-2 cells were significantly increased following the treatment with LPS,which was in a time dependent manner. Compared with LPS group,addition of Rg1 inhibited the increase of PCNA and cyclin D1 in a dose-dependent manner. Conclusion Rg1 regulates the proliferation in activated BV-2 cells through inhibiting the expression of PCNA and cyclin D1.