目的:探讨缺氧复氧损伤环境下Ghrelin对脂肪来源的间充质干细胞(AD-MSCs)的保护作用,以寻求AD-MSCs心肌内移植的有利因素。方法:采用胶原酶消化法分离小鼠AD-MSCs,流式细胞术鉴定其标志。建立缺氧/复氧细胞模型,分3组:①对照组;②缺氧/复氧组(H/R);③H/R+Ghrelin(浓度分别为10-9、10-8、10-7mol/L)干预组。MTT法测定各组细胞增殖,TUNEL法检测细胞凋亡。结果:流式细胞术结果显示AD-MSCs CD44及CD90阳性,CD34、CD45阴性。AD-MSCs MTT分析显示在缺氧环境中,Ghrelin相比于单纯H/R组能够显著促进AD-MSCs的存活与增殖,并抑制其凋亡(P〈0.05)。结论:Ghrelin可以明显提高缺氧复氧环境下AD-MSCs的生存与增殖,抑制缺氧诱导的凋亡发生,有望为心肌梗死的干细胞移植治疗创造新的有利因素。
Objective:To investigate the effect of Ghrelin on adipose tissue derived mesenchymal stem cells(AD-MSCs) under hypoxia-reoxygenation(H/R) injury,and to explore the protective factors for myocardial cell transplantation.Methods:AD-MSCs were isolated from Kunming mice and characterized by flowcytometry.AD-MSCs were divided into(1) Normal control(2) Hypoxia+ Reoxygenation(H/R)(3) H/R+Ghrelin(10-9,10-8,10-7 mol/L,respectively).Cell survival and proliferation were assessed by MTT assays.Apoptosis of AD-MSCs was detected by TUNEL strain.Results:AD-MSCs were positive to the CD44 and CD90.The pretreatment of Ghrelin improved the survival and proliferation of AD-MSCs under H/R condition compared with that of H/R group(P0.05).TUNEL showed Ghrelin inhibited the apoptosis of AD-MSCs after H/R injury(P0.05).Conclusions:Ghrelin has beneficial effects on the survival and proliferation of AD-MSCs under H/R environment and might constitute a hopeful target in stem cell transplantation for ischemic cardiovascular disease.