摘要
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 work demonstrated that bone marrow mesenchymal stem cells （BMSCs） transplantation could improve cardiac function in rats with myocardial infarction. However, the overall efficacy was not satisfactory. The implanted cells presented a low survival rate and newly formed vascular-like structures were sparse in the local infarct tissues. OBJECTIVE： To observe the influence of putative receptor protein related to the angiotensin receptor AT1 endogenous ligand （apelin） on the survival and vascularization potential of BMSCs in hypoxic and ischemic conditions and to investigate relevant mechanisms. METHODS： The bone marrow mesenchymal stem cells were obtained and cultured in vitro with or without apelin-13 （5 pmol/L） stimulation under hypoxic-ischemic condition （serum-free medium, 1% O2） for 24 hours, or cultured under normoxia （complete culture medium, 20% 02） as a negative control during the whole process. All the experimental groups were further co-cultured with human umbilical vein endothelial cells to promote vascular differentiation for another 6 hours. Cell proliferation, apoptosis, and vascular density were assessed and the expression of vascular endothelial growth factor was detected using western blot assay thereafter. RESULTS AND CONCLUSION： Compared with the BMSCs group, BMSCs＋apelin group presented more rapid cell growth, higher proliferation rate, and lower apoptosis percentage under normoxic and hypoxic conditions. In the BMSCs＋apelin group, a larger number of vascular branches formed on matrigel under normoxic and hypoxic-ischemic conditions; and the expression of vascular endothelial growth factor was significantly enhanced. These findings suggest that ape!in could effectively promote BMSCs survival and vascularization under hypoxic-ischemic conditions in vitro. It might function via upregulating the expression of vascular endothelial growth factor. Subject headings： Bone Marrow; Mesenchymal Stem Cells; Receptor, Angiotensin, Type 1 ; Ligands; Cel
出处
《中国组织工程研究》
CAS
北大核心
2017年第1期6-12,共7页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81070125)"抗凋亡与促血管生成miRNA-378干预MSCs治疗心梗后心衰的机制研究"
国家自然科学基金(81270213)"ANGⅡ/AT1/SMAD/CX43通路在心肌干细胞提高心梗大鼠心电生理学稳定性和室颤阈值的作用机制研究"
国家自然科学基金(81670306)"心肌干细胞经IGF-1/STAT/miR-155通路下调AT1R改善心肌梗死后心电生理稳定性机理研究"
广东省科技计划项目(2014A020211002)"LncRNA-Bvht/MESP1/N-cadherin通路调控MSCs向心肌细胞定向分化的机制研究"
广东省科技计划项目(2010B031600032)"抗凋亡与促血管生成miRNA-378干预MSCs治疗心梗后心衰的机制研究"
高校基本科研业务费中山大学青年教师重点培育项目(13ykzd16)"PPAR-γ/TGF-β1/Smad/CX43通路在PPAR-γ干预MSCs治疗心梗后心衰的疗效及机制研究"
广东省医学科研基金(A2016264)"心肌干细胞经由HIF-1α/Apelin/APJ/ACE2通路下调ANGII改善心肌梗死大鼠心电生理学稳定性的机制研究"~~
