摘要
目的:探究骨髓间充质干细胞(BMSCs)在组织工程心脏瓣膜(TEHVs)上的附着状态及脂联素(APN)对BMSCs抗流体剪切应力(FSS)的影响。方法:利用全骨髓法分离培养BMSCs并进行鉴定。首先使用平板流体腔模型,将BMSCs分为对照组、7.5dyne/cm2组、15dyne/cm2组和30dyne/cm2组,接受FSS 24h后观察细胞形态并检测凋亡相关指标。然后将BMSCs细胞分为5组:对照组、15dyne/cm2组、10μmol/L APN+15dyne/cm2组、20μmol/L APN+15dyne/cm2组和30μmol/L APN+15dyne/cm2组,各组在DMEM或APN预处理24h,进而接受FSS 24h后观察细胞形态,检测细胞增殖能力及Bax和Bcl-2的蛋白表达情况。结果:与对照组相比,不同强度的FSS可以显著抑制BMSCs细胞活力、下调Bcl-2表达并且上调Bax表达,并且呈现强度依赖性。与15dyne/cm2组相比,APN预处理可以显著改善细胞形态,增强细胞增殖能力,上调Bcl-2表达,抑制Bax表达。结论:APN预处理能显著减轻BMSCs细胞接受FSS后的损伤,该过程与凋亡相关蛋白Bcl-2及Bax密切相关。
Objective：To investigate whether APN can protect bone marrow mesenchymal stem cell（BMSCs）against fluid shear stress（FSS）.Method：BMSCs were isolated from bone marrow,and identified using specific marker by fluorescence-activated cell sorting analysis s.Four different magnitudes of FSS（0,7.5,15 and 30dynes/cm2）were applied to the BMSCs for 24 hours.BMSCs were pretreated with APN at different concentrations（10,20,and 30μg/ml）for 24 hours.Cell proliferation analysis and western blot analysis were carried out.Result：Compared with the control group（without FSS）,FSS stimulation inhibited cell viability,decreased the expression of Bcl-2and increased the expression of Bax（P〈0.05）in an intensity-dependent manner.BMSCs pretreatment with APN survived better than BMSCs under FSS stimulation,especially at the 30μg/ml concentration（P〈0.05）.Additionally,APN increased the expression of Bcl-2and decreases the expression of Bax（P〈0.05）.Conclusion：APN effectively protects BMSCs against FSS,and the process is closely related to apoptosis related protein Bcl-2and Bax.
出处
《临床心血管病杂志》
CAS
CSCD
北大核心
2016年第5期499-504,共6页
Journal of Clinical Cardiology
基金
国家自然科学基金资助项目(No:81422004
81500263
81470480
81470477
81100137
81100081)
陕西省青年科技新星人才基金(No:2014KJXX-56)
"863计划"青年科学家专题课题(No:2014AA020514
2015AA020919)
国家博士后科学基金(No:2015M572681)
关键词
剪切力
骨髓间充质干细胞
脂联素
组织工程心脏瓣膜
flow shear stress
bone marrow mesenchymal stem cells
adiponectin
tissue-engineered heart valve
