低氧无血清对乳鼠心肌细胞皮肤桥蛋白表达的影响

Effect of hypoxia and serum deprivation on the expression of dermatopontin in cultured neonatal rat cardiomyocytes

目的在体外用低氧无血清条件模拟缺血心肌微环境,探讨心肌细胞皮肤桥蛋白(DPT)表达的变化。方法以低氧无血清处理乳鼠心肌细胞0、6、12和24 h,用实时反转录聚合酶链式反应检测DPT mRNA的表达变化,用Western blot检测DPT的蛋白表达变化。用酶联免疫吸附实验(ELISA)证实低氧无血清处理乳鼠心肌细胞24 hDPT蛋白的表达变化。结果与0 h组相比,低氧无血清处理心肌细胞6、12、24 h DPT的mRNA和蛋白水平均显著升高(P<0.05)。用ELISA法也检测到低氧无血清处理心肌细胞24 h DPT的表达水平高于正常对照组(P<0.05)。结论低氧无血清可促进乳鼠心肌细胞DPT的表达,DPT可能在缺血缺氧引起的心室重构中发挥一定的作用。

Objective To investigate the expression of dermatopontin（DPT） induced by hypoxia and serum deprivation（H/SD） which simulated the microenvironment of ischemic myocardium in cultured neonatal rat cardiomyocytes in vitro.Methods Cultured neonatal rat cardiomyocytes were disposed to H/SD for 0,6,12 and 24 h.The mRNA and protein expression of DPT was determined by real time RT-PCR and western blot,respectively.Using enzyme-linked immunosorbent assay（ELISA） to determine the different expression of DPT between normal cultured cardiomyocytes and cardiomyocytes disposed to H/SD for 24 h.Results Compared the data of 0 h,the mRNA and protein level of DPT were significantly raised in cultured cardiomyocytes which were disposed to H/SD for 6,12 and 24 h（P0.05）.It was confirmed by the method of ELISA that compared with control group,the expression of DPT was higher in cardiomyocytes which were disposed to H/SD for 24 h.ConclusionsThe DPT expression in cultured neonatal rat cardiomyocytes was promoted by H/SD in vitro,which suggested that DPT may be invoved in the ventricular remolding process caused by hypoxia and ischemic in vivo.