培养晚期AD胞质杂交细胞线粒体功能的异常变化  

Mitochondrial functional abnormalities in Alzheimer disease cybrid cell lines at late passage in culture

作　　者：孔令平[1] 麦全安[2] 赵太平[1] 刘妍[1] 汪华侨[3] 

机构地区：[1]广州医科大学从化学院,广东从化510925 [2]广州体育学院 [3]中山大学中山医学院人体解剖学教研室与脑研究室

出　　处：《解剖学研究》2013年第4期256-259,264,共5页Anatomy Research

基　　金：广东省教育厅科技创新项目(2012KJCX0089);广东省建设中医药强省科研课题(20122102);广东省医学科研基金(A2012557);广东省科技计划项目(2010B031600070;2012B031800053);广州市科技计划应用基础研究专项重点项目(2012J410076)

摘　　要：目的探讨AD胞质杂交细胞在培养过程中的功能变化情况,为AD的线粒体发病机制提供实验依据。方法GSH检测试剂盒(比色法)检测细胞匀浆中GSH含量,COX活性检测试剂盒(分光光度计法)测定COX活性,JC-1染色后流式细胞术和激光共聚焦显微镜术检测线粒体膜电位的变化。结果培养晚期AD胞质杂交细胞的GSH含量、COX活性、膜电位水平明显低于培养早期AD胞质杂交细胞,差异有统计学意义(t=8.048,P<0.001;t=12.758,P<0.001和t=2.751,P<0.05),也显著低于培养晚期CTL胞质杂交细胞(t=23.06,P<0.001;t=26.862,P<0.001和t=3.876,P<0.01)。结论AD胞质杂交细胞随着培养时间的延长,其线粒体功能显著降低,这是由于AD患者本身线粒体基因的缺陷所至。Objective To explore the mitoehondrial function changes of AD cybrids and to provide experimental evidence for the mitochondrial pathogenesis of AD. Methods Function of mitochondria in SAD cybrids, CTL cybrids and SYSY cells at ear- ly and late passage were measured by all kinds of methods. GSH contents and COX activity in above cells were respectively detected by chromatometry and spectrophotometer. Mitochondrial membrane potential （MMP, A^m） was investigated using flow eytometry and confocal microscopy after JC-1 staining. Results GSH contents, COX activity and A^m of late passage AD cybrids were sig- nificantly decreased compared with early passage AD cybrids （P 〈 0.001, 0.001 and 0.05 respectively） and late passage CTL cy- brids （P 〈 0.001, 0.001 and 0.01 respectively）. Conclusion Mitochondrial functional abnormalities in AD eybrids worsen with passage in culture. Expression of AD defective mitoehondrial genes results in AD cybrid functional abnormalities.

关 键 词：阿尔茨海默病 AD胞质杂交细胞 氧化应激 线粒体功能 

分 类 号：R749.16[医药卫生—神经病学与精神病学]