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作 者:靳雁斌[1] 司慧远[1] 吴彦瑞 丁学锋[3] 王晓文[3] 范文红[3]
机构地区:[1]空军总医院干部病房五区/神经内科病区,北京100142 [2]解放军第一八八医院检验科,广东潮州521000 [3]军事医学科学院基础医学研究所脑保护与可塑性研究室,北京100850
出 处:《标记免疫分析与临床》2013年第6期427-430,共4页Labeled Immunoassays and Clinical Medicine
基 金:国家自然科学基金青年基金(No.31200822);北京市自然科学基金面上项目(No.5122033)联合资助
摘 要:目的探讨Cpne5蛋白在SH-SY5Y细胞凋亡中表达量的变化。方法显微镜观察细胞形态及流式细胞术检测凋亡率,鉴定A23187诱导SH-SY5Y细胞凋亡及血清剥夺诱导凋亡的模型;免疫印迹法检测两种凋亡过程中Cpne5蛋白表达量的变化。结果 10μmol/L A23187诱导SH-SY5Y细胞24h,细胞出现皱缩、凋亡率增加。A23187诱导0.5h,Cpne5蛋白表达量即出现明显的上升,持续至12h;在24h时Cpne5蛋白表达量回落至诱导前水平。血清剥夺72h的SH-SY5Y细胞24份,凋亡率为23.2±2.1%,与对照组1.1±0.1%比,差异有统计学意义(P<0.01)。免疫印迹结果表明,SH-SY5Y细胞分别经血清剥夺1h、6h、12h、24h、72h对Cpne5蛋白表达量无明显变化。结论 A23187诱导SH-SY5Y细胞凋亡早期Cpne5蛋白表达量有显著变化;提示Cpne5基因可能参与凋亡调控的钙信号通路。Objective To explore the change of Cpne5 protein expression during SH-SY5Y cell apoptosis. Methods The models of A23187 or serum-deprivation inducing SH-SYSY cells apoptosis were used, and the morphology observation and apoptosis rate were detected by flow cytometry. The changes of Cpne5 protein expression during two processes were detected by western blotting. Results After induced by 10μmol/L A23187 for 24 hours, SH-SYSY cells were shrinked and its apoptotic rate was rose. The expression of Cpne5 protein was significantly increased from 0.5 h to 12 h after SH-SY5Y cells treated by A23187, and then reduced to the pre-induced level at 24h. The apoptosis rate of 24 samples of SH-SYSY cells was 23.2 _+2.1% after 72 h ser- umdeprivation ; it was significantly higher than that controls ( 1.1 ±0.1% ). There was no significant change on Cpne5 protein expression when SH-SY5Y cells were treated with serum-deprivation at 1, 6, 12, 24, and 72h. Conclusion Cpne5 protein expression was significantly changed in early apoptosis of SH-SYSY cells when induced by A23187. The Cpne5 gene may be involved in regulation of apoptosis by calcium signaling pathways.
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