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2021 年第 2 期 第 16 卷

血管紧张素-(1-7)及丝裂原活化蛋白激酶激酶抑制剂U0126对血管紧张素Ⅱ诱导的大鼠右心室心肌成纤维细胞增殖的影响

Effects of angiotensin-(1-7) and mitogen activated protein kinase kinase inhibitor U0126 on angiotensin Ⅱ induced proliferation of rat right ventricular cardiac fibroblasts

作者:郭雯1刘文娴1张维君1马涵英2

英文作者:Guo Wen1 Liu Wenxian1 Zhang Weijun1 Ma Hanying2 

单位:1首都医科大学附属北京安贞医院心内科100029;2首都医科大学附属北京安贞医院全科医疗科100029

英文单位:1Department of Cardiology Beijing Anzhen Hospital Capital Medical University Beijing 100029 China; 2General Practice Department Beijing Anzhen Hospital Capital Medical University Beijing 100029 China

关键词:肺动脉高压;右心室重构;转化生长因子β1;Ⅰ型胶原蛋白α1

英文关键词:Pulmonaryarterialhypertension;Rightventricularremodeling;Transforminggrowthfactor-β1;Collagentype1alpha1

  • 摘要:
  • 目的 通过大鼠右心室心肌成纤维细胞(RVCFs)体外培养,探讨血管紧张素(Ang-1-7)及丝裂原活化蛋白激酶激酶抑制剂U0126AngⅡ诱导的RVCFs的影响。方法提取乳大鼠RVCFs,经过抗波形蛋白免疫荧光法鉴定,传代培养至第3代,随机分为对照组、AngⅡ组、Ang-1-7)组(分为低、中、高剂量组)、U0126组(分为低、中、高剂量组)。采用噻唑蓝比色法测定各组细胞增殖情况;蛋白质印迹法检测各组Ⅰ型胶原蛋白α1COL1A1)表达水平;免疫组织化学法测定转化生长因子β1(TGF-β1)表达水平。结果 AngⅡ组RVCFs的细胞增殖率明显高于对照组[(141.7±9.6%比(100.0±0.0%],Ang-(1-7)低、中、高剂量组,U0126低、中、高剂量组RVCFs的细胞增殖率[(118.2±4.9%、(52.1±5.9%、(43.6±2.5%,(43.0±2.3%、(43.3±3.7%、(49.4±4.6%]均显著低于AngⅡ组(均P0.05),且Ang-(1-7)RVCFs增殖的抑制作用呈剂量反应关系。AngⅡ能促进RVCFs COL1A1表达,Ang-(1-7)U0126AngⅡ诱导的COL1A1表达均有抑制作用。TGF-β1免疫组化阳性反应分级示对照组(+)、AngⅡ组(+++)、Ang-(1-7)组(++)、U0126组(+)。结论 Ang-(1-7)U0126可部分抵消AngⅡ刺激导致的RVCFs增殖,抑制TGF-β1COL1A1的蛋白表达。

  • Objective To investigate the effects of angiotensin(Ang)-(1-7) and mitogen activated protein kinase kinase inhibitor U0126 on angiotensin induced rat right ventricular cardiac fibroblasts(RVCFs) cultured in vitro. Methods RVCFs of suckling rats were extracted and identified by anti-vimentin immunofluorescence method. The RVCFs were subcultured to the third generation and randomly divided into control group, Ang group, Ang-(1-7) group including low, medium, high dose groups and U0126 groupincluding low, medium, high dose groups. Methyl thiazolyl trazoliumMTT  was used to detect RVCFs multiplication colorimetric assay. Western blotting was used to measure the expression levels of collagen type 1 alpha 1 COL1A1.  Immunohistochemistry was used to measure the expression levels of transforming growth factor-β1TGF-β1.Results  The cell proliferation rate of RVCFs in Ang group was significantly higher than that of the control group (141.7±9.6)% vs (100.0±0.0)%. The cell proliferation rate of RVCFs in Ang-(1-7) low, medium, high dose groups and U0126 low, middle, high dose groups[(118.2±4.9%,52.1±5.9%,43.6±2.5%,43.0±2.3%,43.3±3.7%,49.4±4.6% were significantly higher than those in Ang group(all P0.05). The inhibitory effect of Ang-(1-7) on the proliferation of RVCFs showed a dose response relationship. Ang could promote the expression of COL1A1 in RVCFs, and Ang-(1-7) and U0126 inhibited the expression of COL1A1 in RVCFs induced by Ang. Immunohistochemical grading of TGF-β1 showed the control group (+), Ang group (+++), Ang-(1-7) group (++), U0126 group (+). Conclusion Ang-(1-7) and U0126 can partially counteract the proliferation of RVCFs induced by Ang and inhibit the protein expression of TGF-β1 and COL1A1.

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