脂肪基质细胞诱导分化为星形胶质细胞过程中Bax/Bcl-2蛋白与细胞凋亡  

Bax/Bcl-2 protein and apoptosis during adipose-derived stromal cell differentiation into astrocytes

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

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

出　　处：《中国组织工程研究》2022年第31期4982-4987,共6页Chinese Journal of Tissue Engineering Research

基　　金：河北省科技计划项目(20567622H),项目负责人:元小冬。

摘　　要：背景:神经退行性疾病是一大类能够导致患者工作和生活能力丧失的重大神经系统疑难疾病,目前对于该病尚无有效的治疗方法,脂肪基质细胞能够在体外诱导分化为神经细胞,有望为该类疾病的治疗提供新的方法。目的:探究脂肪基质细胞诱导分化为星形胶质细胞过程中Bax/Bcl-2蛋白的表达变化,进一步研究诱导过程中是否存在线粒体途径介导的细胞凋亡。方法:应用免疫细胞化学法和Western blot法检测脂肪基质细胞诱导分化为星形胶质细胞48 h及7,14,21 d时星形胶质细胞特异性标志物胶质纤维酸性蛋白,线粒体凋亡相关蛋白Bax、Bcl-2,细胞色素C及Caspase-3的表达,应用透射电子显微镜观察诱导过程中细胞及线粒体的超微结构变化。结果与结论:①在诱导48 h及7,14,21 d时均有胶质纤维酸性蛋白的表达,诱导7 d时胶质纤维酸性蛋白的表达达到高峰期(P<0.05),诱导14 d与诱导7 d表达无明显差异(P>0.05);②随着诱导时间的延长,Bax、细胞色素C及caspase-3的表达水平逐渐升高,至诱导14 d时达到高峰(P<0.05);而Bcl-2的表达水平则逐渐降低,至诱导14 d时达到最低(P<0.05);③透射电镜下可以观察到典型的凋亡小体及线粒体损伤变化;④结果可见,脂肪基质细胞在诱导分化为星形胶质细胞的第7-14天时处于最佳的存活和分化状态。脂肪基质细胞诱导分化为星形胶质细胞的过程中,Bcl-2抑制凋亡的作用明显减弱,Bax的促凋亡作用逐渐增强,细胞色素C通过受损的线粒体释放到胞浆中,从而引发线粒体途径的细胞凋亡。BACKGROUND:Neurodegenerative diseases are a large class of major neurological diseases that can lead to the loss of patients’ability to work and live.At present,there is no effective treatment for this disease.Adipose-derived stromal cells can induce differentiation into nerve cells in vitro,which is expected to provide a new method for the treatment of this kind of diseases.OBJECTIVE:To explore the expression changes of Bax/Bcl-2 protein during the differentiation of adipose-derived stromal cells into astrocytes,and to further investigate whether there is mitochondrial pathway-mediated apoptosis during the induction METHODS:Immunocytochemistry and western blot assay were used to analyze the expression of glial fibrillary acidic protein,Bax,Bcl-2,cytochrome C,and casepase-3 in the process of adipose-derived stromal cell differentiation into astrocytes when induction for 48 hours and 7,14,and 21 days.Transmission electron microscopy was used to observe the ultrastructure of cells and mitochondria.RESULTS AND CONCLUSION:(1)After being induced for 48 hours and 7,14,and 21 days,the expression of glial fibrillary acidic protein reached the peak on day 7(P<0.05).There was no significant difference in glial fibrillary acidic protein expression between 14 days and 7 days after induction(P>0.05).(2)The expression of Bax,cytochrome C,and casepase-3 increased gradually with the induction time extending and reached the peak on day 14(P<0.05).The expression of Bcl-2 was decreased gradually and reached the lowest level on day 14 after induction(P<0.05).(3)Typical apoptotic bodies and mitochondrial damage were observed using transmission electron microscopy.(4)Adipose-derived stromal cells were in the best survival and differentiation state at the 7^(th) to 14^(th) day of differentiation.During adipose-derived stromal cell differentiation into astrocytes,the inhibitory effect of Bcl-2 on apoptosis was significantly weakened,and the pro apoptotic effect of Bax was gradually enhanced.Cytochrome C was released into the cytoplasm

关 键 词：脂肪基质细胞 诱导 分化 星形胶质细胞 线粒体 凋亡 蛋白 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]