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背景:课题组前期研究发现骨髓间充质干细胞移植能够改善大鼠心肌梗死后心功能,但整体效果并不太理想,骨髓间充质干细胞在心肌梗死组织局部的生存力和新生血管密度低下。目的:观察血管紧张素受体AT1相关的受体蛋白内源性配体(apelin)对骨髓间充质干细胞在缺血缺氧条件下的生存和血管形成能力的影响并探讨相关机制。方法:体外培养的骨髓间充质干细胞分为骨髓间充质干细胞组和骨髓间充质干细胞+apelin组,分别于正常条件(完全培养基,体积分数为20%O_2)和缺血缺氧条件(无血清,体积分数为1%O_2)下培养24 h,其中骨髓间充质干细胞+apelin组在培养同时加入apelin-13(5μmol/L)刺激骨髓间充质干细胞。MTS法检测细胞增殖能力,TUNEL法检测细胞凋亡情况。取细胞培养液分别刺激人脐静脉内皮细胞,观察骨髓间充质干细胞血管形成能力。Western blot检测骨髓间充质干细胞中血管内皮生长因子的表达。结果与结论:与骨髓间充质干细胞组相比较,骨髓间充质干细胞+apelin组正常条件和缺血缺氧条件下扩增能力显著增强,凋亡减少,血管管腔样结构明显增多,血管内皮生长因子的表达明显增高。以上结果提示apelin能够增强骨髓间充质干细胞在缺血缺氧环境下的生存和血管形成能力,此效应可能与其上调血管内皮生长因子的表达相关。
Background: Our previous study found that bone marrow mesenchymal stem cell transplantation can improve cardiac function after myocardial infarction in rats, but the overall effect is not very satisfactory. Local viability and neovascular density of bone marrow mesenchymal stem cells in myocardial infarction are low. OBJECTIVE: To observe the effect of apelin, an angiotensin receptor-related receptor protein (AT1) receptor, on the survival and angiogenesis of bone marrow mesenchymal stem cells under hypoxic-ischemic conditions and to explore the underlying mechanisms. METHODS: Bone marrow-derived mesenchymal stem cells (BMSCs) cultured in vitro were divided into BMSC group and BMSC + apelin group, respectively. Under normal conditions (complete medium, volume fraction 20% O 2) and hypoxic- (Serum-free, volume fraction of 1% O2) for 24 h, in which bone marrow mesenchymal stem cells + apelin group while apelin-13 (5μmol / L) stimulation of bone marrow mesenchymal stem cells. MTS assay of cell proliferation, TUNEL assay of apoptosis. Cultured human umbilical vein endothelial cells were stimulated to observe the ability of bone marrow mesenchymal stem cells to form. Western blot was used to detect the expression of vascular endothelial growth factor in bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: Compared with BMSC group, the proliferation ability of BMSC + apelin group under normal and hypoxia-ischemia conditions was significantly increased, the apoptosis decreased, the vascular lumen-like structure was significantly increased, and the vascular The expression of endothelial growth factor was significantly increased. The above results suggest that apelin can enhance the survival and angiogenesis of bone marrow-derived mesenchymal stem cells under hypoxic-hypoxic conditions, which may be related to its upregulation of VEGF expression.