Zhang Y, Yuan R-X, Bao D
Department of Ultrasound, Shanxian Central Hospital, Heze, China.
Eur Rev Med Pharmacol Sci. 2020 May;24(9):4988-4996. doi: 10.26355/eurrev_202005_21190.
To investigate the influence of the Rho/Rho-associated kinase (ROCK) signaling pathway in rats with pulmonary arterial hypertension (PAH) under the intervention with transforming growth factor-beta 1 (TGF-β1).
A total of 30 rats were divided into three groups using a random number table, including control group (healthy rats, n=10), model group (PAH rats, n=10), and TGF group (PAH rats injected with 5 ng/mL TGF-β1 recombinant protein, n=10). The systolic blood pressure, ventricular hypertrophy index, pathological changes in lung tissues, TGF-β1 level, protein, and messenger ribonucleic acid (mRNA) expressions of RhoA and ROCK, as well as concentrations of serum nitric oxide (NO) and endothelin-1 (ET-1) were detected via hemodynamics test, hematoxylin and eosin (HE) staining, immunohistochemical method, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunosorbent assay (ELISA).
The results of hemodynamics test showed that the right ventricular systolic pressure was increased markedly in model group (46.53±8.81) and TGF group (56.79±9.12) compared with that in control group (26.03±4.21) (p<0.05). The mean pulmonary systolic pressure in model group (25.89±1.92) and TGF group (29.41±1.91) was evidently higher than that in control group (15.77±2.71) (p<0.05). According to the results of heart weight measurement, model group (0.5118±0.1635) exhibited a higher ventricular hypertrophy index than control group (0.2908±0.0313) (p<0.05) but a lower ventricular hypertrophy index than TGF group (0.7231±0.1004) (p<0.05). The medial thickness of the pulmonary artery of the rats was observed through the HE staining. It was found that compared with control group, the medial thickness of the pulmonary artery was increased significantly in model group (p<0.05), while it was raised more prominently in TGF group, higher than that in model group, suggesting that TGF-β1 expression can increase the medial thickness of the pulmonary artery. It was manifested in immunohistochemical results that the protein expression of RhoA in the left lung tissues rose notably in model group compared with that in control group (p<0.05), and it was also raised remarkably in TGF group in comparison with that in model group (p<0.05), illustrating that the protein expression of TGF can activate the activity of RhoA and ROCK. The results of RT-PCR indicated that the mRNA expressions of RhoA and ROCK in the left lung tissues were elevated distinctly in model group and TGF group compared with those in control group (p<0.05), and the increases were more apparent in TGF group than those in model group (p<0.05). It was revealed in ELISA results that in comparison with control group, model group, and TGF group had markedly increased concentrations of serum NO and ET-1 (p<0.05), while the rises of serum NO and ET-1 concentrations in TGF group were the most prominent compared with those in model group (p<0.05).
Overexpressed TGF-β1 can activate the RhoA/ROCK signaling pathway, thus promoting the occurrence and development of PAH.
探讨转化生长因子-β1（TGF-β1）干预下Rho/ Rho相关激酶（ROCK）信号通路对肺动脉高压（PAH）大鼠的影响。
采用随机数字表法将30只大鼠分为3组，包括对照组（健康大鼠，n = 10）、模型组（PAH大鼠，n = 10）和TGF组（注射5 ng/mL TGF-β1重组蛋白的PAH大鼠，n = 10）。通过血流动力学检测、苏木精-伊红（HE）染色、免疫组化法、逆转录-聚合酶链反应（RT-PCR）和酶联免疫吸附测定（ELISA）检测收缩压、心室肥厚指数、肺组织病理变化、TGF-β1水平、RhoA和ROCK的蛋白及信使核糖核酸（mRNA）表达，以及血清一氧化氮（NO）和内皮素-1（ET-1）浓度。
血流动力学检测结果显示，与对照组（26.03±4.21）相比，模型组（46.53±8.81）和TGF组（56.79±9.12）的右心室收缩压显著升高（p<0.05）。模型组（25.89±1.92）和TGF组（29.41±1.91）的平均肺动脉收缩压明显高于对照组（15.77±2.71）（p<0.05）。根据心脏重量测量结果，模型组（0.5118±0.1635）的心室肥厚指数高于对照组（0.2908±0.0313）（p<0.05），但低于TGF组（0.7231±0.1004）（p<0.05）。通过HE染色观察大鼠肺动脉中膜厚度。发现与对照组相比，模型组肺动脉中膜厚度显著增加（p<0.05），而TGF组增加更显著，高于模型组，表明TGF-β1表达可增加肺动脉中膜厚度。免疫组化结果显示，与对照组相比，模型组左肺组织中RhoA蛋白表达显著升高（p<0.05），与模型组相比，TGF组也显著升高（p<0.05），说明TGF蛋白表达可激活RhoA和ROCK活性。RT-PCR结果表明，与对照组相比，模型组和TGF组左肺组织中RhoA和ROCK的mRNA表达明显升高（p<0.05），且TGF组升高比模型组更明显（p<0.05）。ELISA结果显示，与对照组相比，模型组和TGF组血清NO和ET-1浓度显著升高（p<0.05），而TGF组血清NO和ET-1浓度升高比模型组最显著（p<0.05）。
TGF-β1过表达可激活RhoA/ROCK信号通路，从而促进PAH的发生发展。
