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

doi:10.16352/j.issn.1001-6325.2023.10.1491

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (10): 1491-1497.doi: 10.16352/j.issn.1001-6325.2023.10.1491

• 研究论文 • 下一篇

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

沈露茜^1#*, 徐学静^2#, 叶逸繁³, 陈卓³, 陈兰³, 申钰琪³, 李红智^3*

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

收稿日期:2023-03-15 修回日期:2023-05-30 出版日期:2023-10-05 发布日期:2023-10-05
通讯作者: *lhz@wmu.edu.cn; imaginary-diva@163.com
作者简介:^#对本文有相同贡献
基金资助:
国家自然科学基金(81971291)

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

SHEN Luxi^1#*, XU Xuejing^2#, YE Yifan³, CHEN Zhuo³, CHEN Lan³, SHEN Yuqi³, LI Hongzhi^3*

1. Department of Internal Neurology, Beijing Friendship Hospital, Capital Medical University, Beijing 100050;
2. Department of Clinical Laboratory, Affiliated Hospital of Southwest Medical University, Luzhou 646000;
3. Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China

Received:2023-03-15 Revised:2023-05-30 Online:2023-10-05 Published:2023-10-05
Contact: *lhz@wmu.edu.cn; imaginary-diva@163.com

摘要/Abstract

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),pS129 α-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。

Abstract: Objective To provide a basis for gene therapy of sporadic Parkinson's disease (PD) caused by mitochondrial complex Ⅰ dysfunction, yeast complex Ⅰ (expressed by internal NADH dehydrogenase,NDI1) was tested to replace human complex Ⅰ with 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 pS129 α-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 complex Ⅰ-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 complex Ⅰ-related functional defects in rotenone-induced differentiated PD cell model, and can alleviate the damage of cytopathology and mitochondrial functions.

Key words: yeast NDI1, recombinant adeno-associated virus (rAAV), rotenone, all-trans retinoic acid, Parkinson's disease

中图分类号:

R741

沈露茜, 徐学静, 叶逸繁, 陈卓, 陈兰, 申钰琪, 李红智. 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[J]. 基础医学与临床, 2023, 43(10): 1491-1497.

SHEN Luxi, XU Xuejing, YE Yifan, CHEN Zhuo, CHEN Lan, SHEN Yuqi, LI Hongzhi. Transduction of yeast NDI1 gene reduces the damages demonstrated by a rotenone-induced differentiated Parkinson's disease cell model[J]. Basic & Clinical Medicine, 2023, 43(10): 1491-1497.

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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

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