心肌成纤维细胞在血管紧张素Ⅱ刺激下表达下调基因的分离和鉴定

Identification of down-regulated genes in cardiac fibroblasts stimulated by angiotensin Ⅱ

目的:对成年大鼠心肌成纤维细胞(CF)受血管紧张素Ⅱ(AngⅡ)刺激后下调基因表达谱进行筛选及分析。方法:以受AngⅡ刺激CF为驱动方,未刺激CF为实验方,进行抑制消减杂交(SSH)建立消减cDNA文库。经斑点杂交筛选文库后将部分阳性克隆测序及进行同源性分析。结果:共获得17条下调表达基因,分别与胞内信号转导、转录抑制、纤维沉积和细胞骨架重构相关,并克隆到4条新基因表达序列标签(EST)。结论:SSH有效地克隆了成年大鼠CF受AngⅡ刺激后下调表达基因,对这些基因的研究将有助于阐明心肌改建的分子机制。

AIM： To identify the down- regulated genes in adult rat cardiac fibroblasts （CF） stimulated with angiotensin Ⅱ （Ang Ⅱ）. METHODS： Suppression subtractive hybridization （SSH） was performed between the CF stimulated by Ang Ⅱ （driver） and unstimulated CF （tester） to generate subtractive cDNA library. The library was screened with dot blots hybridization to further verify the differentially expressed cDNA clones. Partial positive clones were sequenced and BLAST analyzed. RESULTS： Seventeen downregulated genes related to intracellular signal transduction, transcriptional repression, deposition of fibrous matrix and cellular cytoskeletal rearrangement, and 4 new expression sequence tags （EST） were acquired. CONCLUSION： SSH is a powerful technique with high sensitivity for the detection and clone of down - regulated genes expressed in CF induced by Ang Ⅱ , which is helpful to clarify the mechanism of cardiac remodeling.