沉默Fis1基因对METH诱导体外培养SH-SY5Y细胞增殖能力、线粒体膜电位和超微结构的影响  

Effect of silencing Fis1 gene on METH-induced proliferative capacity, mitochondrial membrane potential and ultrastructure of SH-SY5Y cells cultured in vitro

作　　者：黄仕美 牟登峰 王琪 郑丹 齐晓岚[3] 官志忠[3] 于燕妮[1] 楼迪栋 HUANG Shi-mei;MOU Deng-feng;WANG Qi;ZHENG Dan;QI Xiao-lan;GUAN Zhi-zhong;YU Yan-ni;LOU Di-dong(Forensic Pathology Teaching and Research Section,Dept of Legal Medicine in Guizhou Medical University,Guiyang 550004,China;Perinatal Health Care Section,Guiyang Maternal and Child Health Hospital,Guiyang 550004,China;Key Lab of Endemic and Minority Diseases of Ministry of Education,Guizhou Medical University,Key Lab of Medical Molecular Biology of Guizhou Province,Guiyang 550004,China;School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550004,China)

机构地区：[1]贵州医科大学法医学院法医病理学教研室,贵州贵阳550004 [2]贵阳妇幼保健院围产期保健科,贵州贵阳550004 [3]贵州医科大学地方病与少数民族疾病教育部重点实验室,贵州省医学分子生物重点实验室,贵州贵阳550004 [4]贵州中医药大学基础医学院,贵州贵阳550004

出　　处：《中国药理学通报》2019年第12期1725-1730,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81760336);贵阳市科技计划项目(No[2017]30-3)

摘　　要：目的评估线粒体分裂蛋白1(Fis1)在甲基苯丙胺(METH)诱导人神经母细胞瘤细胞(SH-SY5Y细胞)损伤中的作用。方法采用成组设计方法,将体外培养SH-SY5Y细胞分为不同组别:未沉默组、沉默阴性组和沉默组,不同浓度的METH诱导各组SH-SY5Y细胞24 h。利用Western blot检测Fis1蛋白表达水平,使用CCK-8细胞毒性增殖实验分析METH对SH-SY5Y细胞增殖能力的影响,利用线粒体膜电位检测试剂盒(JC-1)检测METH对SH-SY5Y细胞MMP水平的影响,使用透射电镜观察METH对SH-SY5Y细胞线粒体超微结构的影响。结果未沉默组、沉默阴性组和沉默组中,与对照组比较,各组组内随METH诱导SH-SY5Y细胞的浓度升高,Fis1蛋白表达水平升高(P<0.05)、增殖能力减弱(P<0.05)、MMP水平降低(P<0.05);与未沉默组和沉默阴性组相同浓度比较,沉默组SH-SY5Y细胞Fis1蛋白表达水平降低(P<0.05),增殖能力增强(P<0.05),MMP水平升高(P<0.05)。组内与对照组比较,2.0mmol·L^-1 METH诱导未沉默组、沉默阴性组和沉默组,透射电镜观察见线粒体小球状结构增多(P<0.01)。结论Fis1可能在METH诱导体外培养SH-SY5Y细胞损伤中起关键作用。Aim To evaluate the role of fisson 1(Fis1)in methamphetamine(METH)-induced injury of human neuroblastoma(SH-SY5Y)cells cultured in vitro.Methods SH-SY5Y cells cultured in vitro were divided into different groups by the group design method:unsilent groups,silent negative groups and silent groups.Different concentrations of METH induced SH-SY5Y cells in each group for 24 hours.The expression level of Fis1 was detected by Western blot.The effect of METH on the proliferative capacity of SH-SY5Y cells was analyzed by CCK-8 cytotoxicity proliferation assay.The MMP level of METH on SH-SY5Y cells was detected by mitochondrial membrane potential detection kit(JC-1).The effect of METH on the mitochondrial ultrastructure of SH-SY5Y cells was observed by transmission electron microscopy.Results In unsilent group,silent negative group and silent group,the expression level of Fis1 increased(P<0.05)and the proliferative capacity decreased(P<0.05),and the MMP levels decreased(P<0.05)with the increase of the concentration of SH-SY5Y cells induced by METH.Compared with the same concentration in unsilent group and silent negative group,in silent group,the expression level of Fis1 in SH-SY5Y cells decreased(P<0.05),the proliferative capacity increased(P<0.05),and the MMP level increased(P<0.05).Compared with control group,2.0 mmol·L^-1 METH induced unsilent groups,silent negative groups and silent groups,and transmission electron microscopy showed the increase in the mitochondrial small globular structure(P<0.01).Conclusion Fis1 may play a key role in METH-induced injury of SH-SY5Y cells cultured in vitro.

关 键 词：甲基苯丙胺 人神经母细胞瘤细胞 线粒体分裂蛋白1 线粒体膜电位 线粒体超微结构 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学]