贵州医科大学学报

2020, v.45;No.235(04) 387-391+407

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核酸内切酶G在帕金森病大鼠脑组织中的表达及意义
Expression and Significance of Endonuclease-G in Brain Tissue of Rats with Parkinson's Disease

武林;任真奎;吕菊;谢鹏;胡玉梅;吴昌学;禹文峰;
WU Lin;REN Zhenkui;LV Jv;XIE Peng;HU Yumei;WU Changxue;YU Wenfeng;Department of Blood Transfusion,People's Hospital of Southwest Guizhou Autonomous Prefecture;Key Laboratory of Molecular Biology ,Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University;Department of Clinical Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;

摘要(Abstract):

目的:探讨核酸内切酶G(Endo-G)介导的非Caspase依赖性细胞凋亡在帕金森病(PD)发生发展中的作用和分子机制。方法:30只SPF级雄性SD大鼠均分为PD模型组(PD组)和对照组(Ctrl组),PD组大鼠右侧纹状体注射6-羟多巴胺(6-OHDA)诱导建立单侧损毁的PD大鼠模型,Ctrl组大鼠注射生理盐水;采用免疫组化实验观察大鼠黑质内酪氨酸羟化酶(TH)的变化以及核酸内切酶G(Endo-G)在黑质中的分布,计算Endo-G核转移细胞率;采用蛋白印迹实验(Western Bolt)检测Endo-G在黑质线粒体和细胞核中的表达量。结果:Ctrl组TH染色阳性神经元结构完整,PD模型组大鼠损毁侧黑质部位TH染色阳性细胞很难见完整的神经元结构;与Ctrl组比较,PD大鼠模型组损毁侧(右侧)黑质线粒体中Endo-G表达量明显降低(P <0. 05),而细胞核中Endo-G表达量明显升高(P <0. 05); Ctrl组大鼠Endo-G核转移细胞率低于PD组(P <0. 05)。结论:在6-OHDA诱导的PD大鼠模型中,Endo-G自线粒体转位至细胞核,提示非Caspase依赖性细胞凋亡通路可能介导PD大鼠黑质多巴胺能神经元的变形和丢失。
Objective: To explore the role and molecular mechanism of caspase-independent apoptosis mediated by endonuclease G( Endo-G) in the development of Parkinson 's disease( PD).Methods: Thirty male SPF SD rats were divided into PD model group( PD group) and control group( Ctrl group). The rats in PD group were injected with 6-hydroxydopamine( 6-OHDA) into the right striatum to establish PD model, while rats in the Ctrl group were injected with saline.Immunohistochemical( IHC) staining was used to observe the changes of tyrosine hydroxylase( TH)and Endo-G distribution in the substantia nigra. The transfer rate of Endo-G into nuclei was calculated in the substantia nigra. Western blot was used to detect the expression of Endo-G in mitochondria and nuclei in the substantia nigra. Results: The TH-positive neurons had complete neuronal structures in the Ctrl group,while the TH-positive neurons barely had complete neuronal structure in the substantia nigra of the PD model group. When compared with the Ctrl group,mitochondrial Endo-G expression was significantly reduced in PD group( P < 0. 05),while Endo-G expression was significantly increased in the nuclei( P < 0. 05); the transfer rate of Endo-G into nuclei was lower in Ctrl group than in PD group( P < 0. 05). Conclusion: In 6-OHDA-induced PD model,the translocation of Endo-G from mitochondria to the nuclei suggests that the Caspase-independent apoptosis pathway may mediate the deformation and loss of dopaminergic neurons in substantia nigra.

关键词(KeyWords): 帕金森病;羟多巴胺;细胞凋亡;酪氨酸羟化酶;核酸内切酶G;非Caspase依赖性细胞凋亡
parkinson disease(PD);oxidopamine;apoptosis;tyrosine hydroxylase(TH);endonuclease-G(Endo-G);caspase-independent apoptosis

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81360199);; 黔科中引地[2019]4008号;; 贵州省卫生健康委科学技术基金(gzwjkj2019-1-039);; 黔西南州科技局基金(2019-1-10)

作者(Author): 武林;任真奎;吕菊;谢鹏;胡玉梅;吴昌学;禹文峰;
WU Lin;REN Zhenkui;LV Jv;XIE Peng;HU Yumei;WU Changxue;YU Wenfeng;Department of Blood Transfusion,People's Hospital of Southwest Guizhou Autonomous Prefecture;Key Laboratory of Molecular Biology ,Guizhou Medical University;Key Laboratory of Endemic and Ethnic Diseases,Ministry of Education,Guizhou Medical University;Department of Clinical Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;

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DOI: 10.19367/j.cnki.1000-2707.2020.04.003

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