Elabela促进脐带华通胶间充质干细胞向心肌样细胞分化  

Elabela promotes differentiation of Wharton’s jelly-derived mesenchymal stem cells into cardiomyocyte-like cells

作　　者：秦欣宇 张燕[2] 张宁坤 高连如[2] 程涛[1] 王泽 童珊珊[1] 陈宇[1,2] Qin Xinyu;Zhang Yan;Zhang Ningkun;Gao Lianru;Cheng Tao;Wang Ze;Tong Shanshan;Chen Yu(Naval Clinical College of Anhui Medical University,Beijing 100048,China;Department of Cardiovascular Medicine,The Sixth Medical Center of PLA General Hospital,Beijing 100048,China)

机构地区：[1]安徽医科大学海军临床学院,北京市100048 [2]解放军总医院第六医学中心心血管内科,北京市100048

出　　处：《中国组织工程研究》2020年第7期1046-1051,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81370238)，项目负责人：陈宇;北京市自然科学基金(7142156)，项目负责人：陈宇~~

摘　　要：背景:Elabela是近几年发现的新型APJ内源性受体,在成人心血管系统中广泛分布,对心血管疾病也有一定的影响,但是Elabela在干细胞向心肌样细胞诱导分化过程中的作用以及分化过程中对APJ表达量的影响尚未发现相关研究。目的:探讨Elabela对脐带华通胶间充质干细胞向心肌样细胞诱导分化的影响。方法:复苏冻存的脐带华通胶间充质干细胞,当细胞汇合度达到80%-90%时,使用5-氮胞苷使其向心肌样细胞分化,24 h后实验组更换为含Elabela、体积分数为10%胎牛血清的低糖DMEM培养基,对照组更换为含体积分数为10%胎牛血清的低糖DMEM培养基。诱导第7,14,21,28天,观察细胞形态变化,同时收集各组细胞的总RNA和总蛋白,采用实时荧光定量PCR和Western blot技术检测心肌特异性标记物Nkx2.5、cTnT、Connexin 43 mRNA及蛋白表达;采用实时荧光定量PCR和流式细胞技术检测诱导后的心肌样细胞中APJ的表达。结果与结论:①在诱导分化各阶段,实验组心肌特异性标记物Nkx2.5、cTnT、Connexin43 mRNA和蛋白表达量均高于对照组,APJ表达量也高于对照组;②结果表明,Elabela在脐带华通胶间充质干细胞向心肌样细胞分化过程中起一定的促进作用;Elabela作为APJ的另一个激动剂在诱导分化过程中可促进APJ的表达。BACKGROUND:Elabela is a new type of endogenous receptor of APJ discovered in recent years.It is widely distributed in the adult cardiovascular system and has a certain influence on cardiovascular diseases.However,the effect of Elabela on the differentiation of stem cells into cardiomyocytes and the expression of APJ in cardiomyocyte differentiation has not been studied yet.OBJECTIVE:To investigate the effect of Elabela on the differentiation of Wharton’s jelly-derived mesenchymal stem cells into cardiomyocytes.METHODS:The frozen mesenchymal stem cells were resuscitated.5-Azacytidine was used to induce Wharton’s jelly-derived mesenchymal stem cells to differentiate into cardiomyocytes when the cell confluence reached 80%-90%.After 24 hours,the medium was replaced by low-glucose medium containing Elabela and 10% fetal bovine serum in the experimental group,and by low-glucose medium containing 10% fetal bovine serum in the control group.At 7,14,21,and 28 days after induction,cell morphology was observed.The total RNA and total protein of each group were collected.The myocardial specific markers Nkx2.5,cTnT and Connexin 43 mRNA and protein expression levels were detected by real-time fluorescent quantitative PCR and western blot assay.The expression of APJ in the induced cardiomyocytes was detected by real-time fluorescent quantitative PCR and flow cytometry.RESULTS AND CONCLUSION:(1)The expression levels of myocardial specific markers Nkx2.5,cTnT and Connexin 43 mRNA and protein were higher in the experimental group than in the control group in all stages of differentiation,and the expression of APJ was also higher in the experimental group than in the control group.(2)In summary,Elabela plays a certain promoting role in the differentiation of Wharton’s jelly-derived mesenchymal stem cells into oriented cardiomyocytes.Elabela,as another agonist of APJ,can promote the expression of APJ during the induced cell differentiation.

关 键 词：Elabela Apelin13 脐带华通胶 间充质干细胞 5-氮胞苷 APJ 细胞分化 心肌样细胞 

分 类 号：R459.9[医药卫生—治疗学] R394.2[医药卫生—临床医学]