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目的：探讨缺氧诱导因子-1alpha（HIF-1alpha）在转化生长因子beta1（TGF-beta1）促风湿性心脏病心肌细胞胶原合成中的作用。方法：以体外培养风湿性心脏病（风心病）患者经瓣膜置换术后留取组织分离而来的心肌细胞为研究对象，依据加入TGF-beta1 的浓度将前期实验分为四组：0，5，10 及20 (ug/L)，观察TGF-beta1 对风湿性心脏病心肌细胞胶原合成及HIF-1alpha表达的影响；而后实验选取10 ug/L TGF-beta1 为干预浓度，将Scrambled siRNA 或HIF-1-alpha siRNA 转染入细胞内。48 小时后，分别收集各组细胞，采用RT-PCR检测I型胶原的mRNA 水平，Western Blot 技术测定细胞内I 型胶原和HIF-1琢的蛋白表达水平。结果：与0、5 及10 ug/L浓度 TGF-beta1 组相比，5、10 及20 ug/L浓度的TGF-beta1 分别显著地增加了风心病心肌细胞I型胶原及HIF-1-alpha的表达。另外，HIF-1alpha siRNA则明显减少了TGF-beta1 诱导的心肌细胞I型胶原生成。结论：HIF-1-alpha介导了TGF-beta1 对风湿性心脏病心肌细胞I型胶原合成的促进作用。

英文摘要:

Objective:To elucidate the effect of hypoxia inducible factor -1 alpha（HIF-1alpha）on generation of Type I collagen in rheumatic cardiomyocytes induced by transforming growth factor beta 1 (TGF-beta1).Methods:Human rheumatic cardiomyocytes were cultured in vitro, the pre-experiment was divided into 4 groups as follows: 0, 5, 10 and 20 (ug/L), according to the concentration of TGF-beta1. Then, we chose 10 ug/L TGF-beta1, and transfected rheumatic cardiomyocytes with non-specific siRNA or HIF-1alpha siRNA. 48 hours later, the cells were collected. Type I collagen mRNA was detected by RT-PCR, the expression of Type I collagen and HIF-1alpha was determined by Western Blot.Results:Compared with 0, 5, 10 ug/L TGF-beta 1 groups, 5, 10, 20 ug/L TGF-beta1 significantly increased the synthesis of Type I collagen and expression of HIF-1alpha. In addition, HIF-1alpha siRNA markedly decreased generation of Type I collagen induced by TGF-beta 1.Conclusion:HIF-1alpha participates in TGF-beta1 induced synthesis of Type I collagen in rheumatic cardiomyocytes.

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