丙泊酚对新生大鼠脑皮层星形胶质细胞凋亡的影响

Effect of propofol on apoptosis of cortical astrocytes isolated from neonatal rats

目的研究丙泊酚对新生大鼠脑皮层星形胶质细胞凋亡的影响及其机制。方法原代分离和培养AST细胞,用GFAP免疫细胞化学鉴定。不同浓度丙泊酚(0、10、30、90μmol·L-1)作用AST细胞8 h。MMT法检测细胞生存率;Annexin V/PI双染检测细胞凋亡;Western blot法测定线粒体cyt-C漏出;分光光度计法检测caspase-3和caspase-9活性。结果分离培养出原代AST,不同浓度丙泊酚(0、10、30、90μmol·L^(-1))浓度依赖性地降低AST细胞生存率,诱导细胞线粒体cyt-C漏出,上调促凋亡蛋白caspase-3和caspase-9,诱发AST凋亡。结论丙泊酚在体外能诱发新生大鼠脑皮层星形胶质细胞凋亡,其机制可能与丙泊酚诱导线粒体cyt-C释放,激活线粒体凋亡通路有关。

Aim To study the effects of propofol on apoptosis of cortical astrocytes isolated from neonatal rats.Methods Pure astrocytes（AST）were obtained from the cultured cerebral cortical cells and identified by the GFAP stain technology in neonatal rats.AST cells were treated with different concentrations of propofol（0,10, 30, 90 μmol·L^-1）for 8 hours.Cell viability was measured by MTT method, and the apoptosis was detected by Annexin V/PI double staining.Cytochrome C（cyt-C）leakage was detected by Western blot.Caspase-3 and caspase-9 activities were measured by spectrophotometry.Results AST cells were administered with different concentrations of propofol（0,10,30, 90 μmol·L^-1）for 8 hours.It decreased the survival rate in a concentration-dependent manner, induced the leakage of cyt-C in mitochondria, up-regulated the expression of pro-apoptotic protein caspase-3 and caspase-9, and induced AST apoptosis.Conclusion Propofol induces the apoptosis of astrocytes in neonatal rat cortex in vitro, which may be related to the activation of mitochondria apoptosis pathway induced by mitochondrial cyt-C release.