辛伐他汀对脂多糖诱导新生大鼠心肌成纤维细胞NO生成和AT2R蛋白表达抑制的影响

Effects of Simvastatin on LPS induced NO production and AT2R expression in rat cardiac fibroblasts

目的研究辛伐他汀(Sim)对脂多糖(LPS)调控心肌成纤维细胞(CFs)一氧化氮(NO)生成和血管紧张素Ⅱ受体2型(AT2R)蛋白表达的干预效应。方法体外培养新生大鼠CFs,以第1代CFs作为实验对象,采用硝酸还原酶法测定细胞内NO产量,Western blot检测细胞AT2R蛋白的表达,以AT2R/β-actin值对AT2R表达进行半定量。结果与LPS(10mg/L)单独作用组相比,0.01μmol/L和0.1μmol/L Sim与LPS共同作用组CFs的NO生成和AT2R/β-actin值无显著差异;在1μmol/L和10μmol/LSim与LPS共同作用组,NO生成量显著低于LPS单独作用组(均P<0.01),而AT2R/β-actin值显著高于LPS单独作用组(分别有P<0.05和P<0.01)。结论高浓度Sim可拮抗LPS引起的NO生成增加和AT2R蛋白表达下降。

AIM To investigate the effects of Simvastatin （Sim） on lipopolysaccharides （LPS）-induced changes in nitric oxide ac fibroblasts （CFs）. study. After incubating （NO） production and angiotensin Ⅱ type 2 receptor （AT2R） expression in cardi- METHODS First cultures of neonatal rat CFs were employed in the present cells with LPS, the production of intrinsic nitric oxide （NO） was determined using nitrate reductase measurement. The protein levels of AT2R was determined by Western blot analysis. RESULTS No significant difference was observed in NO production and AT2R protein expression （shown as AT2R/β-actin） when CFs were incubated with Sim （0.01 μmolZL） plus LPS （10 mg/L） or Sim （0.1 μmol/L） plus LPS （ 10 mg/L） and incubated with LPS alone. NO production increased significantly when CFs were incubated with 1 μmol/L or 10 μmol/L Sim in the presence of LPS, compared with LPS alone （both P 〈0.01 ）, while AT2R/β-actin decreased significantly in the coincubated groups （P 〈0.05 and P 〈 0.01, respectively）. CONCLUSION High concentrations of Sim antagonize LPS in the induction of NO release and the inhibition of AT2R protein expression.