TRPM7参与血管紧张素Ⅱ介导的心肌成纤维细胞胶原合成

TRPM7 Participate in Angiotensin Ⅱ Induced Cardiac Fibroblasts Collagen Synthesis

目的研究瞬时受体电位通道亚族M7(TRPM7)在血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)介导的心肌成纤维细胞(cardiac fibroblasts,CFs)胶原合成中的作用。方法将分离培养出的SD乳鼠CFs用1μmol/L的AngⅡ干预至不同时间点(0、6、12、24、48h),分别检测CFs上TRPM7蛋白的表达水平,从而找出1μmol/L的AngⅡ诱导CFs上TRPM7蛋白表达的最佳时间点。然后,用腺病毒-TRPM7-shRNA(Ad-TRPM7-shRNA)基因沉默的方法敲低CFs上TRPM7基因的表达,1μmol/L的AngⅡ干预至最佳时间点,检测CFs在TRPM7基因沉默前后胶原合成情况的变化。结果 CFs用1μmol/L的AngⅡ干预至24h后TRPM7蛋白表达量达最高水平;CFs在1μmol/L的AngⅡ干预至24h后,与心脏纤维化相关的胶原蛋白(胶原蛋白Ⅰ、胶原蛋白Ⅲ)合成增多,而在TRPM7基因沉默后这些胶原的合成量则呈明显下降的趋势。结论 TRPM7参与AngⅡ诱导的心肌成纤维细胞胶原合成,从而促进心脏纤维化的发生。

Objective To investigate the role of transient receptor potential melastatin 7 （TRPM7） in angiotensin Ⅱ induced cardiac fibroblasts （CFs） collagen synthesis. Methods The separated cardiac fibroblasts from SD rats were intervened by 1μmol/L AngⅡ to five different time points （0,6,12,24,48h） , and the TRPM7 protein expression levels on CFs membrane was detected respectively ,so as to find out the best time point inducing TRPM7 protein expression of 1 Ixmol/L Ang Ⅱ . Then we knockdown the TRPM7 gene expression on CFs membrane using adenoviruses -TRPM7 -shRNA （Ad -TRPM7 -shRNA） gene silencing , intervened CFs to be the best time point by 1μmol/L Ang Ⅱ and measured collagen expression levels of CFs before and after the TRPM7 gene silencing. Results The protein expression of TRPM7 on CFs membrane reached the highest level when CFs were intervened by 1μmol/L Ang Ⅱ to 24h. After CFs interven- tion,the synthesis of collagen associated with cardiac fibrosis （ such as collagen Ⅰ , collagen Ⅲ ） increase, then decrease obviously after the TRPM7 gene silencing. Conclusion TRPM7 is involved in Ang Ⅱ induced cardiac fibroblasts collagen synthesis, eventually promoting the occurrence of cardiac fibrosis.