柴胡皂苷a抑制PTZ诱导的小鼠海马星形胶质细胞活化

目的:探讨柴胡皂苷a(SSa)对戊四氮(PTZ)诱导的小鼠海马星形胶质细胞活化的抑制作用。方法:分离培养小鼠海马星形胶质细胞,将细胞随机分为对照组、PTZ组、PTZ+0.625 mg/L SSa组和PTZ+1.25 mg/L SSa组。通过免疫荧光染色检测胶质细胞原纤维酸性蛋白(GFAP)的表达来鉴定细胞;用MTT检测评估细胞活力;用流式细胞术检测各组细胞的周期变化;ELISA法检测各组细胞中GFAP和间隙连接蛋白43(Cx43)的表达水平;流式细胞术和Hoechst 33258染色检测各组细胞的凋亡情况。结果:体外原代培养的星形胶质细胞贴壁生长,细胞突起明显。免疫荧光显示星形胶质细胞呈GFAP阳性表达。与对照组比较,PTZ组细胞活力和G_2/M期细胞百分比显著增加(P0.05),GFAP和Cx43的表达水平也显著上调(P0.05);与PTZ组比较,PTZ+0.625 mg/L SSa组和PTZ+1.25 mg/L SSa组细胞活力和G_2/M期细胞百分比均明显下降,GFAP和Cx43的表达水平也降低,但细胞凋亡水平显著增加(P0.05)。结论:SSa能够显著抑制PTZ诱导的海马星形胶质细胞活化,抑制细胞增殖并诱导凋亡。

Saikosaponin a inhibits PTZ-induced activation of hippocampal astrocytes in mice

AIM: To investigate the inhibitory effect of saikosaponin a (SSa) on pentylenetetrazole (PTZ)-induced activation of hippocampal astrocytes in mice. METHODS: Hippocampal astrocytes were isolated and cultured. The cells were randomly divided into control group, PTZ group, PTZ+0.625 mg/L SSa group and PTZ+1.25 mg/L SSa group. The cells were identified by detection of glial fibrillary acidic protein (GFAP). The cell viability was measured by MTT assay. The cell cycle was analyzed by flow cytometry. The expression of GFAP and connexin 43 (Cx43) was mea-sured by ELISA. The level of apoptosis was determined by flow cytometry and Hoechst 33258 staining. RESULTS: The primary hippocampal astrocytes grew by adherent culture, and the processes of the astrocytes were obvious. Immunofluorescence showed positive GFAP expression in the astrocytes. Compared with control group, the viability of the cells and the percentage of the cells in G₂/M phase in PTZ group were significantly increased (P<0.05), and the expression of GFAP and Cx43 was also markedly increased (P<0.05). Compared with PTZ group, the viability of the cells and the percentage of the cells in G₂/M phase were obviously decreased in PTZ+0.625 mg/L SSa group and PTZ+1.25 mg/L SSa group, and the expression of GFAP and Cx43 was also reduced, whereas the percentage of apoptotic cells was significantly increased (P<0.05).CONCLUSION: SSa significantly suppresses PTZ-induced activation of hippocampal astrocytes, inhibits the cell proliferation and induced apoptosis.