摘要
背景:内皮素1作为一种来源于心内膜和动脉血管内皮的旁分泌信号,能够诱导心脏传导细胞的分化成熟。目的:探讨内皮素1在体外诱导P19细胞向心脏传导细胞分化的作用。方法:P19细胞分别与1%二甲基亚砜和100 nmol/L浓度内皮素1孵育,通过Western blot、免疫荧光和qR T-PCR鉴定早期心肌细胞特征性转录因子GATA4、MEF2C和心肌细胞分化标志物MHC-α、cTnT,以及重要的传导细胞转录因子Nkx2.5、Tbx5和分化标志物Cx40、ANP的变化。qRT-PCR鉴定内皮素1特异性受体阻断剂和阻断PI3K-AKT-mTOR信号通路对内皮素1诱导P19细胞分化的影响。结果与结论:(1)内皮素1能够上调P19细胞早期心肌转录因子、心肌结构基因和传导系统特征表型表达;(2)与二甲基亚砜相比,内皮素1能够确实诱导P19细胞向心脏传导细胞分化;(3)内皮素1主要通过ETA通路上调Nkx2.5和Cx40表达水平从而诱导P19细胞向传导细胞分化,ETB通路可能还参与了一条负反馈调节通路;(4)内皮素1能够通过PI3K-AKT-mT OR信号通路调控Nkx2.5表达进而影响传导细胞分化。
BACKGROUND:Endothelin-1,as a paracrine cytokine derived from endocardium and vascular endothelial cells,can promote the differentiation and maturation of cardiac conduction cells.OBJECTIVE:To explore the effect of endothelin-1 on P19 cells differentiating into cardiac conduction cells in vitro.METHODS:P19 cells were cultured with 1%dimethyl sulfoxide or 100 nmol/L endothelin-1.Afterwards,the changes of early cardiomyocyte transcription factors MEF2C and GATA4,differentiation markers MHC-αand cTnT,important conduction cell transcription factors Nkx2.5 and Tbx5,and differentiation markers Cx40 and ANP were identified by western blot assay,immunofluorescence and real-time PCR.Specific antagonists of endothelin-1 receptors,and blocking PI3K-AKT-mTOR signaling pathway on the differentiation of P19 cells induced by endothelin-1 were evaluated by real-time PCR.RESULTS AND CONCLUSION:Endothelin-1 significantly up-regulated the levels of early cardiac transcription factors,cardiomyocyte structure factors and characteristic conduction system markers.Compared with dimethyl sulfoxide,endothelin-1 was prone to enhance the differentiation of cardiac conduction cells derived from P19 cells.Up-regulation of Nkx2.5 and Cx40 by endothelin-1 was mainly attributed to the ETA signaling pathway.ETB signaling pathway may be also involved in a negative feedback regulation pathway.The PI3K-AKT-mTOR signaling pathway can play an important role in the differentiation from P19 cells to cardiac conduction cells triggered by endothelin-1.
作者
陈浩
张武
王浩
张恒一
徐志伟
Chen Hao;Zhang Wu;Wang Hao;Zhang Heng-yi;Xu Zhi-wei(Department of Cardiothoracic Surgery,Shanghai Children’s Medical Center,Shanghai Jiao Tong University School of Medicine,Shanghai 200127,China;Institute for Hematology,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第12期1915-1921,共7页
Chinese Journal of Tissue Engineering Research
