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

作　　者：吕菊 谢鹏 任真奎 胡玉梅 张春林 吴昌学[1,2] 焦玲[5] 禹文峰[1,2] 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,Guiyang 550004,Guizhou,China;Education Ministry Key Laboratory of Endemic and Minority Diseases of Guizhou Medical University,Guiyang 550004,Guizhou China;Department of Laboratory,People's Hospital of Southwest Guizhou Autonomous Prefecture,Xingyi 562400,Guizhou,China;College of Basic Medical,Guizhou Medical University,Guiyang 550025,Guizhou,China;Department of Neurology,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学分子生物学重点实验室,贵州贵阳550004 [2]贵州医科大学地方病与少数民族疾病教育部重点实验室,贵州贵阳550004 [3]黔西南州人民医院检验科,贵州兴义562400 [4]贵州医科大学基础医学院,贵州贵阳550025 [5]贵州医科大学附院神经内科,贵州贵阳550004

出　　处：《贵州医科大学学报》2020年第3期255-259,共5页Journal of Guizhou Medical University

基　　金：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)。

摘　　要：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 125μmol/L),and the cell viability was measured by CCK8 to select the optimal concentration of 6-OHDA.The control cells and Seipin-knockdown 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 75μmol/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：羟多巴胺 PC12细胞 帕金森病 细胞凋亡 内质网应激 Seipin蛋白 

分 类 号：R742.5[医药卫生—神经病学与精神病学] R-332[医药卫生—临床医学]