脂肪基质细胞诱导为星形胶质细胞过程中线粒体融合/分裂蛋白的变化及与凋亡的关系  

Changes of mitochondrial fusion/fission proteins and their relationship with apoptosis during adipose-derived stromal cells differentiation into astrocytes

作　　者：张丽丽 欧亚[1] 张萍淑 元小冬 ZHANG Lili;OU Ya;ZHANG Pingshu;YUAN Xiaodong(Department of Neurology,Kailuan General Hospital,Tangshan 063000,China;Hebei Key Laboratory of Neurobiology,Tangshan 063000,China)

机构地区：[1]开滦总医院神经内科,唐山063000 [2]河北省神经生物机能重点实验室,唐山063000

出　　处：《生命的化学》2023年第8期1317-1322,共6页Chemistry of Life

基　　金：河北省科技计划项目(20567622H);唐山市科学技术研究与发展计划(201302100)。

摘　　要：将脂肪基质细胞(adipose-derived stromal cells,ADSC)在体外诱导分化为星形胶质细胞(astrocytes,AS),按诱导时间分为诱导48 h组、7 d组、14 d组以及21 d组,分别应用免疫细胞化学法检测胶质纤维酸性蛋白(GFAP)、线粒体融合蛋白2(mitofusin 2,Mfn2)、线粒体分裂蛋白1(fission 1,Fis1)、半胱氨酸天冬氨酸特异性蛋白酶-3(Caspase-3)的表达,并应用流式细胞仪AnnexinⅤ/PI双染法检测各组细胞的存活状态,探讨了ADSC诱导分化为AS过程中线粒体融合、分裂相关蛋白的变化及其与凋亡的关系。结果发现,ADSC诱导分化为AS过程中,Mfn2的阳性表达率随诱导时间延长逐渐降低,至诱导14 d时达到最低;Fis1和Caspase-3的阳性表达率随诱导时间延长逐渐增加,至诱导14 d时达到高峰;细胞的存活率逐渐下降,凋亡率逐渐增加。结果提示,在ADSC诱导为AS的过程中,随着诱导时间的延长,Mfn2表达不足及Fis1的过表达可能会导致细胞凋亡相关蛋白Caspase-3的表达增加,导致细胞凋亡的发生。Adipose-derived stromal cells(ADSC)were induced into astrocytes(AS)in vitro and divided into 48 h,7 d,14 d,and 21 d groups according to the induction time.Immunocytochemical method was used to detect the expression of glial fibrillary acidic protein(GFAP),mitochondrial fusion protein 2(Mfn2),mitochondrial fission 1(Fis1),cysteine aspartate specific protease-3(Caspase-3).The survival status of cells in each group was detected by flow cytometry using AnnexinⅤ/PI double staining method to investigate the changes of mitochondrial morphological regulatory proteins(mitochondrial fusion,mitochondrial fission proteins)during ADSC induced differentiation into AS and their relationship with apoptosis.The results showed that during ADSC induced differentiation into AS,the positive expression rate of Mfn2 gradually decreased with the prolongation of induction time,and reached the lowest level at the 14th day of induction;the positive expression rates of Fis1 and Caspase-3 gradually increased with the prolongation of induction time,reaching a peak at 14 days of induction;The survival rate of cells gradually decreases,while the apoptosis rate gradually increases.The results suggest that during the induction of ADSC into AS,the insufficient expression of Mfn2 and the overexpression of Fis1 may lead to an increase in the expression of apoptosis related protein Caspase-3,leading to the occurrence of cell apoptosis,as the induction time prolongs.

关 键 词：脂肪基质细胞 星形胶质细胞 线粒体融合 线粒体分裂 凋亡 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]