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胚胎干细胞(ESC)具有无限增殖和分化为体内3个胚层来源的各种类型组织细胞的潜能,经过体外诱导能够分化为心肌细胞,亦称为胚胎干细胞分化心肌细胞(ESCM).本研究探讨了ESC诱导分化心肌细胞过程中血管紧张素Ⅱ受体(ATR)的亚型AT1R和AT2R的表达特征.10-4mol/L维生素C体外诱导小鼠R1胚胎干细胞分化为自发搏动的心肌细胞,用免疫荧光法检测分化后的细胞表达心肌细胞特异性标志物α辅肌动蛋白.小鼠胚胎干细胞在诱导分化为心肌细胞以后,逆转录聚合酶链反应(RT-PCR)和实时定量RT-PCR(Real-timeRT-PCR)方法检测到ESCM表达AT1R,并且呈时间依赖性逐渐增加的特点,在第14d达到高峰.Western印记法检测AT1R表达特征与RT-PCR结果相符.Western印记法的结果显示,血管紧张素Ⅱ(10-6mol/L)可作为AT1R激动剂激活AT1R,并使其下游的细胞外信号调节激酶(ERK1/2)磷酸化水平上调,预孵育AT1R抑制剂Losartan(10-6mol/L),此作用被抑制.RT-PCR方法显示,与新生小鼠心室肌细胞相比,ESCM的AT2R表达水平较低.
Embryonic stem cells (ESCs) have the potential to proliferate indefinitely and differentiate into various types of tissue cells derived from three germ layers in vivo and are capable of differentiating into cardiomyocytes, also known as embryonic stem cell-derived cardiomyocytes (ESCM), after in vitro induction ESC induces the expression of AT1R and AT2R in the subtype of angiotensin Ⅱ receptor (ATR) during the differentiation of cardiomyocytes.10-4mol / L vitamin C induces the differentiation of mouse R1 embryonic stem cells into spontaneous beating cardiomyocytes in vitro, Fluorescence method was used to detect differentially expressed cardiomyocyte-specific marker α-actinin.Mouse embryonic stem cells were induced to differentiate into cardiomyocytes by reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR The expression of AT1R in ESCM was detected by real-time RT-PCR (RT-PCR) and the expression of AT1R was detected by RT-PCR and Western blotting showed that the expression of AT1R in ESCM was increased in a time-dependent manner. Angiotensin II (10-6 mol / L) can activate AT1R as an AT1R agonist and upregulate the phosphorylation of extracellular signal-regulated kinase (ERK1 / 2) downstream of it. Pretreatment with AT1R inhibitor Losartan (1 0-6mol / L), this effect was inhibited.RT-PCR method showed that, compared with neonatal mouse ventricular myocytes, the expression of AT2R low level of ESCM.