Apelin干预骨髓间充质干细胞移植治疗改善心肌梗死后心功能  

Apelin combined with bone marrow mesenchymal stem cells transplantation improves cardiac function after myocardial infarction

作　　者：侯婧瑛[1] 龙会宝[1] 陈旭翔[1] 汪蕾[1] 郭天柱[1] 郑韶欣[1] 钟婷婷[1] 伍权华[1] 吴浩[1] 王彤[1] Hou Jing-ying;Long Hui-bao;Chen Xu-xiang;Wang Lei;Guo Tian-zhu;Zheng Shao-xin;Zhong Ting-ting;Wu Quan-hua;Wu Hao;Wang Tong(Department of Emergency,Sun Yat-Sen Memorial Hospital,Sun Yat-Sen University,Guangzhou 510120,Guangdong Province,China)

机构地区：[1]中山大学孙逸仙纪念医院急诊科,广东省广州市510120

出　　处：《中国组织工程研究》2018年第25期3937-3943,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81700242)"lnc RNA-H19/miR-199a-5p/VEGF通路在apelin促进MSCs生存和血管再生改善心梗后心衰中的作用及机制研究";广东省科技计划项目(2017A020215176)"Apelin激活PI3K/AKT经lnc RNA-H19/miR-199a-5p/VEGF途径促进MSCs生存和血管再生改善心肌梗死后心力衰竭机制研究";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";广东省医学科研基金(A2017001)"Apelin经PI3K/AKT/mi R-199a-5p/VEGF通路促进骨髓间充质干细胞生存和血管再生改善心肌梗死后心力衰竭机制研究"~~

摘　　要：背景:课题组前期研究发现血管紧张素受体AT1相关的受体蛋白内源性配体(apelin)能够促进骨髓间充质干细胞(mesenchymal stem cells,MSCs)在体外缺血缺氧条件下的生存和血管再生。目的:采用体内实验探讨apelin联合MSCs移植治疗对心肌梗死后心功能的影响及相关机制。方法:开胸结扎50只C57BL/6小鼠左前降支冠状动脉并随机分为5组:MSCs组、MSCs+apelin组、sh-APJ MSCs+apelin组、apelin组和PBS组。模型建立2周后,对动物进行第2次开胸手术,MSCs组于局部梗死心肌内注射单纯的MSCs,MSCs+apelin组和sh-APJ MSCs+apelin组分别注射MSCs以及sh-APJ MSCs并且均连续腹腔内注射2周apelin-13,PBS组心肌内注射不含MSCs的PBS。各组治疗前及治疗2周后检测心功能。实验结束后,摘取心脏检测左心室心肌组织梗死边缘区的MSCs生存和血管再生情况以及相关因子的表达。结果与结论:(1)与治疗前比较,治疗2周后MSCs组、MSCs+apelin组、sh-APJ MSCs+apelin组、apelin组左室射血分数明显增加(P<0.01),左室舒张末期内径、左室收缩末期内径显著减小(P<0.01);其中MSCs+apelin组以上指标改善最显著(P<0.01);(2)与MSCs组和sh-APJ MSCs+apelin组相比,MSCs+apelin组梗死边缘区PKH26阳性细胞和CD31阳性细胞的平均光密度值显著增高,且PKH26和CD31双阳性细胞占PKH26阳性细胞的比例显著增加(P<0.01);(3)与MSCs组和sh-APJ MSCs+apelin组相比较,MSCs+apelin组APJ、AKT、p AKT以及VEGFA表达显著增加,而MSCs组和sh-APJ MSCs+apelin组在上述各项指标表达方面差异均无显著性意义(P>0.05);(4)以上提示apelin能够通过与其受体APJ结合改善MSCs在心肌梗死局部组织中的生存和血管再生,从而进一步提高心肌梗死后心功能,此效应与VEGFA表达上调相关,PI3K/AKT的激活可能在其中起到一定的调控作用。BACKGROUND:Our previous work demonstrated that apelin could promote bone marrow mesenchymal stem cells(BMSCs)survival and vascularization under hypoxic-ischemic conditions in vitro.OBJECTIVE:To explore the effect of apelin combined with BMSCs transplantation on cardiac function after myocardial infarction and the relevant mechanisms.METHODS:Fifty C57BL/6 mice with myocardial infarction were induced by the left anterior descending coronary artery ligation.Animal models were then randomized into five groups:BMSCs,BMSCs+apelin,sh-APJ BMSCs+apelin,apelin and PBS groups.Two weeks after modeling,the mice in the former four groups were subjected to secondary thoracotomy,BMSCs group was given intramyocardial injection of sole BMSCs.BMSCs+apelin and sh-APJ BMSCs+apelin groups were given intramyocardial injection of BMSCs and sh-APJ BMSCs respectively,in combination with intraperitoneal injection of apelin-13 for 2 weeks consecutively.Mice in the PBS group received intramyocardial injection of PBS without BMSCs.Cardiac function of mice was evaluated before and 2 weeks after treatment.MSCs survival and vascularization in the infarct border zone were examined,and the expression of relevant factors was detected thereafter.RESULTS AND CONCLUSION:(1)At 2 weeks after treatment,the cardiac function of mice was obviously improved in the former four groups in contrast with the baseline,especially in the BMSCs+apelin group;Left ventricular ejection fraction was significantly increased(P<0.01),and left ventricular end diastolic diameter and left ventricular end systolic diameter were significantly reduced(P<0.01).(2)Compared with BMSCs and sh-APJ BMSCs+apelin groups,the average optical density of PKH26-and CD31-positive BMSCs in the infarct border zone was significantly enhanced in the BMSCs+apelin group.Moreover,the percentage of CD31 and PKH26 double positive cells to the PKH26-positive BMSCs was considerably increased(P<0.01).(3)The expression levels of APJ,AKT,pAKT and VEGFA were significantly increased in the BMSCs+apelin group

关 键 词：血管紧张素受体AT1相关的受体蛋白内源性配体 骨髓间充质干细胞 心肌梗死 蛋白激酶B 血管内皮生长因子 干细胞 国家自然科学基金 骨髓 间质干细胞移植 心肌梗塞 血管内皮生长因子A 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]