贵州医科大学学报

2020, v.45;No.234(03) 255-259

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Seipin敲低对内质网应激通路诱导细胞凋亡的影响
Effect of Seipin Knockdown on Apoptosis Induced by Endoplasmic Reticulum Stress Pathway

吕菊;谢鹏;任真奎;胡玉梅;张春林;吴昌学;焦玲;禹文峰;
LV Jv;XIE Peng;REN Zhenkui;HU Yumei;ZHANG Chunlin;WU Changxue;JIAO Ling;YU Wenfeng;Key Laboratory of Molecular Biology of Guizhou Medical University;Education Ministry Key Laboratory of Endemic and Minority Diseases of Guizhou Medical University;Department of Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;College of Basic Medical,Guizhou Medical University;Department of Neurology,the Affiliated Hospital of Guizhou Medical University;

摘要(Abstract):

目的:探讨Seipin敲低对肾上腺嗜铬细胞瘤(PC12)细胞凋亡的影响及其相关机制,初步探索Seipin敲低在帕金森病(PD)模型中的可能调控机制。方法:对细胞给予不同浓度的6-羟多巴胺(6-OHDA,0、25、50、75、100及125μmol/L)处理,用CCK8法检测细胞活性筛选6-OHDA最佳造模浓度;选取阴性对照细胞和Seipin敲低细胞分别用6-OHDA诱导体外PD模型作为阴性模型组和Seipin模型组,同时设阴性正常组和Seipin正常组; 4组细胞培养18 h时,采用蛋白质免疫印迹法(Western blot)检测内质网应激(ERS)蛋白(GRP78和CHOP)和凋亡相关蛋白Bax水平。结果:CCK8法检测细胞活性,6-OHDA最适作用浓度为75μmol/L; Western blot结果显示,与阴性正常组比较,阴性模型组、Seipin模型组GRP78、CHOP及Bax蛋白水平明显增加,差异有统计学意义(P <0. 05);而与阴性模型组相比,Seipin模型组GRP78、CHOP及Bax蛋白水平明显增加,差异有统计学意义(P <0. 05)。结论:敲低Seipin诱导细胞凋亡可能涉及ERS蛋白的增加,敲低Seipin后导致6-OHDA诱导的细胞凋亡更加严重。
Objective: To investigate the effect of Seipin knockdown on apoptosis of PC12 cells and its related mechanisms,and to explore the possible regulatory mechanism of seipin knockdown in Parkinson's disease( PD) model. Methods: PC12 cells were treated with different concentrations of 6-hydroxydopamine( 6-OHDA,0,25,50,75,100,and 125 μmol/L),and the cell viability was measured by CCK8 to select the optimal concentration of 6-OHDA. The control cells and Seipinknockdown cells were used to induce PD model in vitro with 6-OHDA as the negative model group and seipin model group respectively,while the negative normal group and Seipin normal group also set up.The cells of 4 groups were cultured for 18 h,Western blot were used to detect the levels of endoplasmic reticulum stress proteins( GRP78 and CHOP) and apoptosis-related protein-Bax. Results: The optimal concentration of 6-OHDA was 75 μmol/L by CCK8. Western blot results showed that the levels of GRP78,CHOP and Bax proteins in the negative model group and the Seipin model group increased significantly( P < 0. 05) compared to the negative normal group. The levels of GRP78,CHOP and Bax proteins in the Seipin model group were significantly increased( P < 0. 05) compared to the negative model group. Conclusion: The apoptosis induced by knockdown of seipin may involve the increase of endoplasmic reticulum stress. Knockdown of Seipin leads to more serious apoptosis induced by 6-OHDA.

关键词(KeyWords): 羟多巴胺;PC12细胞;帕金森病;细胞凋亡;内质网应激;Seipin蛋白
hydroxydopamine;pc12 cells;parkinson's disease(PD);endoplasmic reticulum stress;apoptosis;seipin protein

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金(81360199);; 黔科中引地[(2019)4008];; 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): 吕菊;谢鹏;任真奎;胡玉梅;张春林;吴昌学;焦玲;禹文峰;
LV Jv;XIE Peng;REN Zhenkui;HU Yumei;ZHANG Chunlin;WU Changxue;JIAO Ling;YU Wenfeng;Key Laboratory of Molecular Biology of Guizhou Medical University;Education Ministry Key Laboratory of Endemic and Minority Diseases of Guizhou Medical University;Department of Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture;College of Basic Medical,Guizhou Medical University;Department of Neurology,the Affiliated Hospital of Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2020.03.002

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