NOX4-siRNA转染对AngⅡ诱导的心肌成纤维细胞增殖和胶原合成影响  

EFFECT OF NOX4-SIRNA TRANSFECTION ON CARDIAC FIBROBLAST PROLIFERATION AND COLLAGEN SYNTHESIS INDUCED BY ANG Ⅱ

作　　者：何兆辉 王志谦 王国良 郑先杰 滕伟 HE Zhaohui;WANG Zhiqian;WANG Guoliang;ZHENG Xianjie;TENG Wei(Department of Cardiology,First Affiliated Hospital of Henan University,Kaifeng 475000,China)

机构地区：[1]河南大学第一附属医院心血管内科,河南开封475000

出　　处：《青岛大学学报（医学版）》2021年第5期742-745,共4页Journal of Qingdao University(Medical Sciences)

基　　金：河南省医学科技攻关计划项目(2018020314)。

摘　　要：目的探讨转染NADPH氧化酶4(NOX4)小干扰RNA(siRNA)对血管紧张素Ⅱ(AngⅡ)诱导的心肌成纤维细胞(CFs)增殖和胶原合成的影响及其机制。方法分离乳鼠CFs,将CFs分为对照组(正常培养)、AngⅡ组(加入10^(-6)mol/L AngⅡ)、AngⅡ+siNOX4组(转染NOX4-siRNA后加入10^(-6)mol/L AngⅡ)和AngⅡ+siNC组(转染NC-siRNA后加入10^(-6)mol/L AngⅡ)。用免疫印迹法检测各组NOX4、CollagenⅠ、CollagenⅢ、p-p38MAPK、p38MAPK的表达,CCK-8法检测细胞增殖。结果与对照组相比,AngⅡ刺激后NOX4、CollagenⅠ、CollagenⅢ和p-p38MAPK蛋白的表达水平和细胞增殖活力均明显升高(F=140.134~349.556,q=17.963~25.587,P<0.05);转染NOX4-siRNA后AngⅡ刺激引起上述变化明显减弱(q=15.839~36.742,P<0.05)。结论沉默NOX4可通过阻碍p38MAPK信号通路活化抑制AngⅡ诱导的CFs增殖和胶原合成。Objective To investigate the effect of transfection with NADPH oxidase 4(NOX4) small interfering RNA(siRNA) on the proliferation of cardiac fibroblasts(CFs) and collagen synthesis induced by angiotensin Ⅱ(Ang Ⅱ) and its mechanism. Methods CFs were isolated from neonatal mice and divided into control group(normal culture), Ang Ⅱ group(adding 10^(-6) mol/L Ang Ⅱ), Ang Ⅱ+siNox4 group(adding 10^(-6) mol/L Ang Ⅱ after transfection with NOX4 siRNA), and Ang Ⅱ+siNC group(adding 10^(-6) mol/L Ang Ⅱ after transfection with NC siRNA). Western blot was used to determine the expression of NOX4, collagen Ⅰ, collagen Ⅲ, phosphorylated p38 mitogen-activated protein kinase(p-p38 MAPK), and p38 mitogen-activated protein kinase(p38 MAPK). Cell Counting Kit-8 assay was used to determine the cell proliferation. Results Compared with the control group, the Ang Ⅱ group had significantly increased expression levels of NOX4, Collagen Ⅰ, Collagen Ⅲ, and p-p38 MAPK proteins and cell proliferation activity(F=140.134-349.556,q=17.963-25.587,P<0.05);the changes induced by Ang Ⅱ were significantly decreased after transfection with NOX4 siRNA(q=15.839-36.742,P<0.05). Conclusion Silencing NOX4 can inhibit the proliferation of CFs and collagen synthesis induced by Ang Ⅱ by blocking activation of the p38 MAPK signaling pathway.

关 键 词：肌细胞 心脏 肌成纤维细胞 血管紧张素Ⅱ NADPH氧化酶4 细胞增殖 胶原Ⅲ型 

分 类 号：R329.411[医药卫生—人体解剖和组织胚胎学] R345.4[医药卫生—基础医学]