氯喹抑制体外培养星形胶质细胞的激活

Chloroquine inhibits the activation of astrocyte in vitro

目的研究氯喹对体外培养大鼠海马星形胶质细胞激活的抑制作用,为癫痫的治疗提供实验依据。方法分离新生SD大鼠海马,体外培养星形胶质细胞,经纯化鉴定后分为:对照组、戊四氮(PTZ)组、氯喹干预组(25、50和75 mg/L),经相应处理后,分别用MTT法、免疫荧光、Western blot测定星形胶质细胞数量及活性、星形胶质纤维酸性蛋白(GFAP)及CyclinD1的表达量。结果与对照组比较,PTZ可激活星形胶质细胞的增殖,使GFAP、CyclinD1的表达量增加(P<0.05);与PTZ组比较,氯喹阻滞了PTZ激活的星形胶质细胞的增殖(P<0.05);氯喹可抑制PTZ激活星形胶质细胞异常增加的GFAP的表达;氯喹可抑制PTZ激活星形胶质细胞的CyclinD1的表达量(P<0.05);与对照组相比,3种结果均显示75 mg/L氯喹对体外培养星形胶质细胞激活的抑制作用较强,并可维持其在正常范围。结论氯喹具有抑制PTZ激活体外培养星形胶质细胞的作用,其可能通过抑制星形胶质细胞的增殖来发挥抗癫痫作用。

Objective To investigate the effects of Chliroquine on Pentylenetetrazole（PTZ）-activated hippocampal astrocytes of rats.Methods eonatal rat（24h） hippocampal astrocytes were obtained and divided into control group,PTZ-activated group and Chliroquine groups with Chliroquine treatment at 25,50,75 mg/L.The cell proliferation,expression of glial fibrillary acidic protein（GFAP） and CyclinD1 after the treatments were detected with MTT assay,immunofluorescence cytochemistry and western blot respectively.Results N Results In comparison with control group,PTZ could induce the activation of astrocytes and increase the expressions of GFAP and CyclinD1（P0.05）,In comparison with PTZ-activation group,Chliroquine treatment resulted in significant inhibition of the cell proliferation,GFAP and CyclinD1 in the PTZ-activated astrocytes（P0.05）.Chliroquine at 75 mg/L showed the strongest inhibitory effects against astrocyte activation while maintained nearly normal level of astrocyte activation in comparison with the control group.Conclusions Chliroquine could inhibit PTZ-induced activation of astrocytes,suggesting that chliroquine may function as potential medicine for epilepsy.