The studies of the functional significance of regenerating gene (Reg) in failing hearts.

1.为了证明Reg基因在大鼠心脏中的激活谱，我们建立了两种急性心力衰竭模型——冠状动脉结扎引起的心肌梗死和主动脉收缩引起的压力过载。real-time RT-PCR检测心脏组织中Reg mRNA及Reg受体mRNA的表达水平。Reg mRNA表达水平与急性心脏应激严重程度呈正相关。Reg-1 mRNA主要在心房活化，reg -受体mRNA在受损心室强烈表达。总之，我们的研究结果首次证明了Reg/Reg受体系统在受损心脏中的存在。我们发现REG-1蛋白在死于心肌梗死的尸检患者的心脏中表达。免疫组织化学染色显示REG蛋白在心肌细胞胞浆中呈细颗粒状。心肌细胞以外的组织成分似乎不参与REG蛋白的分泌。使用拉伸装置（ST-140, Strex Inc .），将培养的心肌细胞（H9c2）在柔性硅橡胶腔上拉伸10%或15%，拉伸1、3和6小时。拉伸心肌细胞未检测到Reg基因的表达。由于本研究中观察到的ANP和BNP mRNA的表达谱与先前报道的文献不同，因此需要进一步研究Reg基因在拉伸心肌细胞中的表达。

1.To demonstrate the activation profiles of Reg gene expression in the rat heart, we generated two models of acute heart failure - myocardial infarction by coronary artery ligation and pressure-overload by aortic constriction. The expression levels of Reg mRNA and Reg-receptor mRNA in the heart tissue were quantified by a real-time RT-PCR. There was a positive correlation between the level of Reg mRNA expression and the severity of acute heart stress. Reg-1 mRNA activation was observed mainly in the atria, and Reg-receptor mRNA was expressed intensely in damaged ventricles. In conclusion, our results demonstrate for the first time the presence of the Reg/Reg-receptor system in damaged hearts.2.We found REG-1 protein expression in the human hearts obtained from autopsied patients who died of myocardial infarction. REG protein was immunohistochemically stained in a fine granular pattern in the cytoplasm of cardiomyocytes. Tissue components other than cardiomyocytes appeared not to be involved in the secretion of REG protein.3.Cultured cardiomyocytes (H9c2) were stretched 10% or 15% on flexible silicon rubber chamber for 1, 3, and 6 hours, using stretching apparatus (ST-140, Strex Inc). No Reg gene expression was detected in the stretched cardiomyocytes. Because the expression profiles of ANP and BNP mRNA observed in the present study were different from previously reported literatures, further studies were needed to elucidate the Reg gene expression in the stretched cardiomyocytes.