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为了探讨体外高糖诱发内皮细胞损伤而继发性对人脐带间充质干细胞(human umbilical cord-derived mesenchymal stem cell,h UC-MSC)增殖和凋亡的影响及其机制,该研究采用酶消化法分离培养人脐静脉内皮细胞(human umbilical vein endothelial cell,h UVEC),制备不同糖浓度(5,30 mmol/L)培养h UVECs 24,48,72 h条件培养基(conditioned medium,CM);内皮细胞条件培养基(h UVEC-CM)培养h UC-MSC 3 d,实时细胞监测系统检测h UC-MSC的增殖;Annexin V/PI双染流式细胞术检测细胞的凋亡情况;Western blot法检测细胞凋亡相关蛋白Bcl-2、Bax和Cl-Caspase3的表达。结果显示,条件培养基培养h UC-MSC 3 d后,与对照组相比,h UVEC-CM(HG)组h UC-MSC的增殖活力明显降低,凋亡率显著升高,凋亡相关蛋白Bcl-2的表达明显减少,Bax及Cl-Caspase3的表达明显增加(P<0.05)。以上表明,h UVEC-CM(HG)能抑制h UC-MSC的增殖,并通过调节凋亡相关蛋白Bcl-2、Bax、Cl-Caspase3的表达诱导h UC-MSC凋亡。
To investigate the effect and mechanism of secondary on the proliferation and apoptosis of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) induced by high glucose in vitro, we used enzymatic digestion Methods Human umbilical vein endothelial cells (hUVECs) were isolated and cultured to prepare conditioned medium (CM) of hUVECs at different concentrations of 5 and 30 mmol / L for 24, 48 and 72 h. H UC-MSC was cultured with hUVEC-CM for 3 days. The proliferation of h UC-MSCs was detected by real-time cell monitoring system. The apoptosis of hUC-MSCs was detected by Annexin V / PI double staining flow cytometry. Method to detect the expression of apoptosis-related proteins Bcl-2, Bax and Cl-Caspase3. The results showed that the proliferative activity of h UC-MSCs in hUVEC-CM (HG) group was significantly decreased and the apoptosis rate was significantly increased after 3 h culture of h UC-MSCs in conditioned medium compared with the control group. The apoptosis-related protein Bcl-2 expression was significantly decreased, Bax and Cl-Caspase3 expression was significantly increased (P <0.05). The above results indicate that hUVEC-CM (HG) can inhibit hUC-MSC proliferation and induce apoptosis of hUC-MSCs by regulating the expression of Bcl-2, Bax and Cl-Caspase3.