心肌成纤维细胞在血管紧张素Ⅱ作用中活性氧水平及p22^(phox)的表达

Angiotensin Ⅱ increases ROS production in cardiac fibroblasts by inducing p22～(phox) over-expression

目的探讨血管紧张素Ⅱ(AngⅡ)干预心肌成纤维细胞时活性氧的改变及可能的产生途径,为防治心室重塑发展提供思路。方法培养出生2～3d大鼠心肌成纤维细胞,分为正常对照组,AngⅡ(10-7mol/L)组,APO(100μmol/L)组,APO+AngⅡ(APO100μmol/L培养1h后加入终浓度10-7mol/LAngⅡ)组。干预48h后应用荧光显微镜观测活性氧水平;免疫组织化学法检测p22phox蛋白表达。结果AngⅡ组活性氧荧光最强,APO+AngⅡ组明显减弱,对照组和APO组最弱。组化染色结果显示对照组和APO组几乎未见p22phox蛋白阳性表达;AngⅡ组细胞普遍阳性表达,显著高于其他3组;APO+AngⅡ组稍强于对照组和APO组,但无显著性差异。结论AngⅡ可能通过增强p22phox蛋白表达,导致NADPH氧化酶途径产生活性氧增多。

Objective To investigate the changes in the reactive oxygen species （ROS） in rat cardiac fibroblasts exposed to angiotensin Ⅱ （Ang Ⅱ） treatment and explore the possible pathways that mediate ROS production. Methods In vitro cultured fetal rat cardiac fibroblasts treated with apocynin （APO, 100 μmol/L）, Ang Ⅱ （10.7 mol/L）, or APO＋Ang Ⅱ （10^-7 mol/L Ang Ⅱ was added 1 h after 100 μmol/L APO）, and the ROS levels and p22^phox expression in the cells were detected using fluorescent microscope and immunohistochemistry, respectively. Results Compared with the normal control cells, Ang Ⅱ treatment of the cardiac fibroblasts resulted in significantly increased ROS production, the effect of which was inhibited by the application of APO. p22^phox expression was hardly detected by immunohistochemistry in the control cells, but over-expressed in AngⅡ-treated cells. APO substantially decreased the over-expression of p22^phox induced by Ang Ⅱ. Conclusion Ang Ⅱ increases ROS production in fetal rat cardiac fibroblasts probably by inducing p22^phox over-expression.