论文部分内容阅读
目的:探讨NADPH氧化酶4(NOX4)在小鼠心脏纤维母细胞转化为心肌纤维母细胞中的作用及其可能机制。方法:分离并培养原代C57BL/6小鼠心脏纤维母细胞,分为空白对照组、TGF-β1组(10μg/L TGFβ-1作用24 h)、TGFβ-1+DPI组(10μmol/L DPI预处理2 h后,TGFβ-1作用24 h)和TGFβ-1+siRNA组(NOX4siRNA预处理6 h后,TGFβ-1作用24 h)。α平滑肌肌动蛋白(SMα-actin)和NOX4的mRNA含量采用Real-time PCR检测。SMα-actin蛋白水平采用western blotting检测。H2O2含量采用Amplex Red法检测。结果:NOX4 siRNA能显著抑制细胞NOX4 mRNA的表达。TGFβ-1组细胞内NOX4、SMα-actin mRNA表达、SMα-actin蛋白含量和H2O2水平均高于空白对照组;而DPI和siRNA则明显抑制了TGF-β1诱导的NOX4和SMα-actin表达上调,同时减少了H2O2的产生。结论:NOX4及其催化的ROS部分介导了TGFβ-1诱导的心脏纤维母细胞向心肌纤维母细胞的分化。
Objective: To investigate the role of NADPH oxidase 4 (NOX4) in the transformation of mouse cardiac fibroblasts into cardiac fibroblasts and its possible mechanism. Methods: Primary cardiac fibroblasts from C57BL / 6 mice were isolated and cultured and divided into blank control group, TGF-β1 group (10 μg / L TGFβ-1 for 24 h), TGFβ-1 + DPI group After pretreatment for 2 h, TGFβ-1 was treated for 24 h) and TGFβ-1 + siRNA group (TGFβ-1 for 24 h after NOX4 siRNA pretreatment for 6 h). α-smooth muscle actin (SMα-actin) and NOX4 mRNA content using Real-time PCR detection. The SMα-actin protein level was detected by western blotting. H2O2 content using Amplex Red test. Results: NOX4 siRNA significantly inhibited the expression of NOX4 mRNA. The expression of NOX4 and SMα-actin mRNA, the content of SMα-actin and the level of H2O2 in TGFβ-1 group were higher than those in blank control group, while DPI and siRNA significantly inhibited the upregulation of NOX4 and SMα-actin induced by TGF-β1, While reducing the production of H2O2. Conclusion: NOX4 and its catalytic ROS partially mediate TGFβ-1-induced cardiac fibroblast differentiation into cardiac fibroblasts.