导入酵母NDI1基因减少鱼藤酮诱导的分化型帕金森病细胞模型的损伤  

Transduction of yeast NDI1 gene reduces the damages demonstrated by a rotenone-induced differentiated Parkinson′s disease cell model

作　　者：沈露茜 徐学静 叶逸繁 陈卓 陈兰 申钰琪 李红智[3] SHEN Luxi;XU Xuejing;YE Yifan;CHEN Zhuo;CHEN Lan;SHEN Yuqi;LI Hongzhi(Department of Internal Neurology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050;Department of Clinical Laboratory,Affiliated Hospital of Southwest Medical University,Luzhou 646000;Key Laboratory of Laboratory Medicine,Ministry of Education,School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]首都医科大学附属北京友谊医院神经内科,北京100050 [2]西南医科大学附属医院医学检验部,四川泸州646000 [3]温州医科大学检验医学与生命科学学院教育部检验医学重点实验室,浙江温州325035

出　　处：《基础医学与临床》2023年第10期1491-1497,共7页Basic and Clinical Medicine

基　　金：国家自然科学基金(81971291)。

摘　　要：目的探讨导入酵母NDI1基因能否替代人功能缺陷的线粒体复合体Ⅰ,为复合体Ⅰ功能障碍所致散发型帕金森病(PD)的基因治疗提供研究基础。方法将表达酵母NDI1的重组腺相关病毒(rAAV-NDI1)感染鱼藤酮诱导的分化型帕金森病细胞模型。设DMSO+空载、鱼藤酮+空载、鱼藤酮+NDI1共3组。用Western blot、免疫荧光染色、氧耗测定、ATP含量测定、ROS测定等方法检测细胞病理学、线粒体功能方面指标。结果鱼藤酮+NDI1组较鱼藤酮+空载组,细胞形态明显改善,细胞存活率显著上升(P<0.05),pS129α-突触核蛋白、全细胞自噬显著下降(P<0.05,P<0.001);复合体Ⅰ依赖性氧耗显著升高(P<0.01),细胞总ATP合成、线粒体氧化磷酸化偶联的ATP合成显著上升(P<0.01),线粒体ROS、线粒体自噬显著降低(P<0.01,P<0.001)。结论导入酵母NDI1基因可以在鱼藤酮诱导的分化型帕金森病细胞模型中,替代性弥补复合体Ⅰ的功能缺陷,对细胞病理学、线粒体功能的损伤具有改善作用。Objective To provide a basis for gene therapy of sporadic Parkinson′s disease(PD)caused by mitochondrial complexⅠdysfunction,yeast complexⅠ(expressed by internal NADH dehydrogenase,NDI1)was tested to replace human complexⅠwith functional defects.Methods The recombinant adeno-associated virus(rAAV-NDI1)infected the rotenone-induced differentiated cell model of PD.Three groups(DMSO+vector,rotenone+vector,rotenone+NDI1)were designed.The cytopathology and mitochondrial functions were examined by the methods of Western blot,immunofluorescence staining,measurement of oxygen consumption,ATP content and ROS,etc.Results Compared with rotenone+vector group,the rotenone+NDI1 group showed significantly improved cell morphology,increase in cell survival(P<0.05),and decrease in level of pS129α-synuclein and of whole-cell autophagy(P<0.05,P<0.001).Compared with rotenone+vector group,the rotenone+NDI1 group also displayed significant increase of complexⅠ-dependent oxygen consumption(P<0.01),significant increase in total cellular ATP synthesis and mitochondrial oxidative phosphorylation-coupled ATP synthesis(P<0.01),significant decrease in the level of mitochondrial ROS and mitochondrial mitophagy(P<0.01,P<0.001).Conclusions Yeast NDI1 can replace and compensate complexⅠ-related functional defects in rotenone-induced differentiated PD cell model,and can alleviate the damage of cytopathology and mitochondrial functions.

关 键 词：酵母NDI1 重组腺相关病毒(rAAV) 鱼藤酮 全反式维甲酸 帕金森病 

分 类 号：R741[医药卫生—神经病学与精神病学]