文献类型：期刊文献

中文题名：当归补血汤经TGF-β1/Smad2通路对Ang Ⅱ诱导肥大心肌细胞的保护机制研究

英文题名：Protective effect of Danggui Buxue Tang on TGF-β1/Smad2 pathway in Ang Ⅱ-induced hypertrophic cardiomyocytes

作者：张永花[1];何建新[1];颜春鲁[1];金华[1];徐厚谦[1]

第一作者：张永花

机构：[1]甘肃中医药大学

第一机构：甘肃中医药大学

年份：2018

卷号：34

期号：2

起止页码：9

中文期刊名：中药药理与临床

外文期刊名：Pharmacology and Clinics of Chinese Materia Medica

收录：北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；

基金：甘肃自然科学基金;基金号码:145RJZA170

语种：中文

中文关键词：当归补血汤;血管紧张素Ⅱ;心肌细胞;TGF-β1/Smad2信号通路

外文关键词：dangguibuxuetang;angiotensin Ⅱ;cardiomyocytes;TGF-β1/Smad2 signaling pathway

摘要：目的:研究当归补血汤对血管紧张素Ⅱ(AngⅡ)诱导H9C2肥大心肌细胞的保护作用及机制,探讨其与转化生长因子β1(TGF-β1)及下游分信号Smad2信号通路的关系。方法:体外培养细胞,用α-actin抗体免疫组化法鉴定心肌细胞;采用10-7mol/L AngⅡ诱导H9C2心肌细胞肥大,通过检测心肌细胞蛋白含量及心钠素(ANF)mRNA表达以验证心肌细胞肥大模型;在模型建立成功的基础上利用逆转录聚合酶链反应RT-PCR法测定不同浓度(5%、10%、15%、20%)当归补血汤含药血清对肥大心肌细胞ANF mRNA表达的影响,选取最佳干预浓度;将心肌细胞分组为空白对照组、10-6mol/L AngⅡ模型组、10-7mol/L AngⅡ模型组和10%含药血清组,镜下观察各组细胞形态,并利用蛋白免疫印迹法(Western blot)检测各组TGFβ1/Smad2信号通路相关蛋白TGFβ1、Smad2表达与活化情况。结果:鉴定细胞符合心肌细胞特征;与空白对照组相比,10-7mol/L AngⅡ模型组心肌细胞蛋白含量与ANFmRNA表达均显著增加,说明10-7mol/L AngⅡ诱导H9C2心肌细胞肥大模型建立;10%、15%当归补血汤含药血清浓度组较模型组ANF mRNA表达显著减少,10%当归补血汤含药血清组ANF表达降低更为明显;形态学观察,AngⅡ模型组心肌细胞密度和形态增大,漂浮细胞增多;10-6mol/L、10-7mol/L AngⅡ模型组较空白对照组TGF-β1、Smad2蛋白表达均显著增多,10%当归补血汤含药血清组较10-6mol/L、10-7mol/L模型组TGF-β1、Smad2蛋白表达均显著减少。结论:当归补血汤含药血清具有抗心肌细胞肥大的作用,其保护机制可能与调控心肌细胞TGF-β1/Smad2信号通路有关。
Objective： To investigate the protective effect and mechanism of Danggui Buxue Decoction on cardiomyocyte induced by angiotensin II（ Ang II） in H9 C2 hypertrophied cardiomyocytes,and to investigate its relationship with transforming growth factor β1（ TGF-β1） and downstream signal Smad2 signaling pathway. Methods： Cells were cultured in vitro and myocardial cells were identified byα-actin antibody immunohistochemistry. H9 C2 cardiomyocyte hypertrophy was induced by 10-7 mol/L Ang II,and the cardiomyocyte hypertrophy model was verified by detecting the myocardial protein content and the expression of atrial natriuretic peptide（ ANF） mRNA. Based on the successful establishment of the model,effects of different concentrations（ 5%,10%,15%,20%） of serum containing Danggui Buxue Decoction on the expression of ANF mRNA in hypertrophic cardiomyocytes were determined by reverse transcription-polymerase chain reaction（ RT-PCR）. The best intervention concentration was chosed. Cardiomyocytes were divided into the control group,10-6 mol/L Ang II model group,10-7 mol/L Ang II model group and 10% serum containing drug group. The cell morphology in each group was observed under microscope and Western blot was performed. Expressions and activations of TGFβ1 and Smad2 were detected in each group of TGFβ1/Smad2 signaling pathway related proteins. Results： Characteristics of identified cells were consistent with them of cardiomyocytes. Compared with the control group,the protein content of the cardiomyocytes and the expression of ANF mRNA were increased in the 10-7 mol/L Ang II model group（ P 〈 0. 05）,indicating that H9 C2 cardiomyocyte hypertrophy model induced by 10-7 mol/L Ang II was set up. Compared with the model group,10% and 15% serum containing Danggui Buxue Decoction decreased（ P 〈 0. 05）. The expression of ANF in 10% serum containing Dang Gui Bu Xue Decoction group more obviously decreased. Compared with 10-7 mol/L Ang II model group,the expression of ANF mRNA was not statistically significant（ P 〉 0. 05）. Morphological observation showed that the density and morphology of myocardial cells in Ang II model group increased,and floating cells increased. Protein expressions of TGF-β1 and Smad2 in the 10-6 mol/L and 10-7 mol/L Ang II model group were higher than those in the control group（ P 〈 0. 05）. Compared with 10-6 mol/L and 10-7 mol/L model group,expressions of TGF-β1 and Smad2 protein in 10%serum containing Dang Gui Bu Xue Decoction group decreased（ P 〈 0. 05）. Conclusion： Serum containing Dangguibuxue decoction has the anti-cardiomyocyte hypertrophy,and its protective mechanism may be related to the regulation of cardiac myocyte TGF-β1/-Smad2 signaling pathway.