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[摘要] 目的 探討转染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10-6mol/L AngⅡ）、AngⅡ+siNOX4组（转染NOX4-siRNA后加入10-6mol/L AngⅡ）和AngⅡ+siNC组（转染NC-siRNA后加入10-6mol/L AngⅡ）。用免疫印迹法检测各组NOX4、CollagenⅠ、Collagen Ⅲ、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、Collagen Ⅲ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。
[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
[中图分类号] R329.411;R345.4
[文献标志码] A
[文章编号] 2096-5532（2021）05-0742-04
doi：10.11712/jms.2096-5532.2021.57.170
[开放科学（资源服务）标识码（OSID）]
[网络出版] https：//kns.cnki.net/kcms/detail/37.1517.R.20211029.1735.007.html;2021-11-01 16：47：01
EFFECT OF NOX4-SIRNA TRANSFECTION ON CARDIAC FIBROBLAST PROLIFERATION AND COLLAGEN SYNTHESIS INDUCED BY ANG Ⅱ
HE Zhaohui， WANG Zhiqian， WANG Guoliang， ZHENG Xianjie， TENG Wei
（Department of Car-
diology， First Affiliated Hospital of Henan University， Kaifeng 475000， China）
[ABSTRACT] 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-p38MAPK）， and p38 mitogen-activated protein kinase （p38MAPK）. 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-p38MAPK 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 p38MAPK signaling pathway.
方法 分离乳鼠CFs，将CFs分为对照组（正常培养）、AngⅡ组（加入10-6mol/L AngⅡ）、AngⅡ+siNOX4组（转染NOX4-siRNA后加入10-6mol/L AngⅡ）和AngⅡ+siNC组（转染NC-siRNA后加入10-6mol/L AngⅡ）。用免疫印迹法检测各组NOX4、CollagenⅠ、Collagen Ⅲ、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、Collagen Ⅲ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。
[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
[中图分类号] R329.411;R345.4
[文献标志码] A
[文章编号] 2096-5532（2021）05-0742-04
doi：10.11712/jms.2096-5532.2021.57.170
[开放科学（资源服务）标识码（OSID）]
[网络出版] https：//kns.cnki.net/kcms/detail/37.1517.R.20211029.1735.007.html;2021-11-01 16：47：01
EFFECT OF NOX4-SIRNA TRANSFECTION ON CARDIAC FIBROBLAST PROLIFERATION AND COLLAGEN SYNTHESIS INDUCED BY ANG Ⅱ
HE Zhaohui， WANG Zhiqian， WANG Guoliang， ZHENG Xianjie， TENG Wei
（Department of Car-
diology， First Affiliated Hospital of Henan University， Kaifeng 475000， China）
[ABSTRACT] 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-p38MAPK）， and p38 mitogen-activated protein kinase （p38MAPK）. 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-p38MAPK 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 p38MAPK signaling pathway.