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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 the adhesion of bone marrow mesenchymal stem cells (BMSCs) to tissue-engineered heart valves (TEHVs) and the effect of adiponectin (APN) on the anti-fluid shear stress (FSS) of BMSCs. Methods: BMSCs were isolated and identified by whole bone marrow method. First, BMSCs were divided into control group, 7.5 dyne / cm2 group, 15 dyne / cm2 group and 30 dyne / cm2 group by using flat fluid chamber model. After receiving FSS for 24 hours, the cell morphology was observed and apoptosis related indexes were detected. The BMSCs were divided into 5 groups: control group, 15dyne / cm2 group, 10μmol / L APN + 15dyne / cm2 group, 20μmol / L APN + 15dyne / cm2 group and 30μmol / L APN + 15dyne / cm2 group. The cells were pretreated with DMEM or APN for 24 h, then were treated with FSS for 24 h. The cell proliferation and the protein expression of Bax and Bcl-2 were detected. Results: Compared with the control group, different concentrations of FSS could significantly inhibit the viability of BMSCs, down-regulate the expression of Bcl-2 and up-regulate the expression of Bax in a dose-dependent manner. Compared with 15dyne / cm2 group, APN pretreatment can significantly improve cell morphology, enhance cell proliferation, up-regulate Bcl-2 expression and inhibit Bax expression. CONCLUSION: APN pretreatment can significantly reduce the damage of BMSCs after receiving FSS, which is closely related to the apoptosis-related proteins Bcl-2 and Bax.