摘要
背景:研究表明,脂肪组织来源干细胞能够有效改善心肌梗死后心功能,是一个非常有前景的心肌细胞再生来源。目前大部分研究是在正常氧体积分数下探讨脂肪干细胞介导的心肌保护作用。目的:观察低氧环境培养条件下脂肪干细胞的旁分泌及抗凋亡等保护作用。方法:分离、培养大鼠脂肪细胞以及乳鼠心肌细胞,在正常氧体积分数下和低氧体积分数下(10%)共培养大鼠脂肪干细胞及过氧化氢损伤的乳鼠心肌细胞,通过酶联免疫吸附法检测细胞沉淀中还原型谷胱甘肽与氧化型谷胱甘肽比值(GSH/GSSG)以及培养基上清中血管内皮生长因子、胰岛素样生长因子、碱性成纤维细胞生长因子水平,Western blot检测细胞沉淀中凋亡蛋白Bax和Bcl-2的表达。结果与结论:(1)过氧化氢损伤乳鼠心肌细胞后,还原型谷胱甘肽与氧化型谷胱甘肽比值、血管内皮生长因子、胰岛素样生长因子、碱性成纤维细胞生长因子水平下降;(2)在低氧状态下,损伤的心肌细胞与脂肪干细胞共培养能够使还原型谷胱甘肽与氧化型谷胱甘肽比值升高,促进脂肪干细胞旁分泌细胞因子,而且使凋亡蛋白Bax表达下降,Bcl-2表达上升;(3)结果表明,低氧状态促进脂肪干细胞旁分泌作用增强,从而起到抗凋亡、保护心肌的作用。
BACKGROUND:Adipose-derived stem cells(ADSCs)represent one of the promising cell sources for myocardial regeneration due to their easy accessibility and efficacy in the improvement of cardiac function following myocardial infarction.However,previously reported studies on the underlying mechanism of ADSCs-mediated cardioprotective effect mainly focused on the ADSCs cultured at room air.OBJECTIVE:To test the paracrine actions and anti-apoptotic effect of ADSCs under hypoxic conditions.METHODS:After isolation and culture,neonatal rat myocardial cells were injured by hydrogen peroxide and co-cultured with rat ADSCs under normoxia and hypoxia(10%O2)conditions.Ratio of reduced glutathione to oxidized glutathione(GSH/GSSG)in the cell pellet and levels of vascular endothelial growth factor(VEGF),insulin-like growth factor 1(IGF-1),and basic fibroblast growth factor(bFGF)were tested by ELISA.Expression of apoptotic proteins Bax and Bcl-2 were determined by western blot.RESULTS AND CONCLUSION:GSH/GSSG,VEGF,IGF-1,and bFGF were decreased in neonatal rat myocardial cells injured by hydrogen peroxide.ADSCs significantly attenuated hydrogen peroxide-induced myocardial apoptosis by increasing the ratio of GSH/GSSG and the secretion of VEGF,IGF-1 and bFGF.ADSCs also down-regulated Bax expression and up-regulated Bcl-2 expression.To conclude,hypoxic conditions can enhance the anti-apoptosis and cardioprotective effects of ADSCs through the paracrine mechanism.
作者
高宇平
林媛媛
李学文
范春辉
杨帆
郝大洁
葛文佳
Gao Yu-ping;Lin Yuan-yuan;Li Xue-wen;Fan Chun-hui;Yang Fan;Hao Da-jie;Ge Wen-jia(Department of Cardiovascular Medicine Taiyuan 030032,Shanxi Province,China;Department of Equipments Taiyuan 030032,Shanxi Province,China;Interventional Therapy Center Taiyuan 030032,Shanxi Province,China;Office of Academic Affairs,Shanxi Dayi Hospital,Taiyuan 030032,Shanxi Province,China;Department of Emergency,Taiyuan Central Hospital,Taiyuan 030009,Shanxi Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第9期1344-1349,共6页
Chinese Journal of Tissue Engineering Research
基金
山西省重点研发计划(国际合作)项目(2014081050-2)
关键词
干细胞
脂肪组织
肌细胞
心脏
共同培养技术
细胞低氧
组织工程
Stem Cells
Adipose Tissue
Myocytes,Cardiac
Coculture Techniques
Cell Hypoxia
Tissue Engineering
