白藜芦醇通过TGF-β1/Smad3信号通路抑制心肌成纤维细胞中AngⅡ诱导的Ⅰ/Ⅲ型胶原合成  

Resveratrol inhibits the expression of typeⅠ and type Ⅲ collagen induced by AngⅡin cardiac fibroblasts through regulating the TGF-β1/Smad3 signaling pathway

作　　者：卓小桢[1] 刘懿[2] 李娜[2] 尹晶晶[2] 习文 刘军辉[1,2] ZHUO Xiao-zhen;LIU Yi;LI Na;YIN Jing-jing;XI Wen;LIU Jun-hui(Department of Cardiology;Department of Clinical Laboratory,the First Affiliated Hospital of Xi＇an Jiaotong University,Xi＇an 710061,China)

机构地区：[1]西安交通大学第一附属医院心血管内科,陕西西安710061 [2]西安交通大学第一附属医院检验科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2018年第4期525-529,557,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81500219;81400302);陕西省科学技术研究发展计划项目(No.2016SF-217)~~

摘　　要：目的探讨心肌成纤维细胞(CFs)中白藜芦醇(Res)对血管紧张素Ⅱ(AngⅡ)诱导的胶原释放的抑制效果,及TGF-β/Smads信号通路在其中的作用。方法提取大鼠原代CFs,免疫荧光鉴定细胞内vimentin抗原的表达,以确定细胞种类;2~8代大鼠CFs,给予不同条件干预,Real-time PCR检测Ⅰ型及Ⅲ型胶原、TGF-β1mRNA的表达水平;Western blot检测Ⅰ型及Ⅲ型胶原、TGF-β1、Smad-3、P-Smad-3蛋白的表达水平;ELISA方法检测培养基上清中Ⅰ型胶原、TGF-β1蛋白的分泌水平。结果大鼠CFs提取成功,vimentin抗原表达明显;与普通培养基相比较,AngⅡ干预显著促进CFs中Ⅰ型及Ⅲ型胶原、TGF-β1mRNA和蛋白表达(P<0.05),Res显著抑制AngⅡ干预诱导的Ⅰ型及Ⅲ型胶原、TGF-β1mRNA和蛋白表达(P<0.05);与普通培养基相比较,AngⅡ干预显著促进CFs中Smad-3磷酸化(P<0.05),Res显著抑制AngⅡ诱导的Smad-3磷酸化(P<0.05),各种干预对Smad-3蛋白表达水平无显著影响;AngⅡ培养基上清中Ⅰ型胶原蛋白、TGF-β1浓度显著高于普通培养基(P<0.05),AngⅡ培养基中添加Res显著抑制AngⅡ诱导的Ⅰ型胶原、TGF-β1蛋白的分泌(P<0.05)。结论白藜芦醇可能通过TGF-β1/Smad3信号通路抑制CFs中AngⅡ诱导的Ⅰ/Ⅲ型胶原合成。Objective To explore the inhibitory effect of resveratrol on hypertension-related myocardial fibrosis and the key role of TNF-β/Smads signaling pathway in the anti-fibrosis of resveratrol.Methods The expression of vimentin in the primary rat CFs was evaluated by immunofluorescence to determine the cell type.CFs were treated in different conditions,the mRNA expression levels of typeⅠ and typeⅢ collagen and TGF-β1 were detected by Real-time PCR;the protein expression levels of type Ⅰ and type Ⅲ collagen,TGF-β1,Smad-3 and PSmad-3 were detected by Western blot.The secretion levels of typeⅠcollagen and TGF-β1 in CFs supernatant were measured by ELISA assay.Results Rat CFs were successfully extracted,and vimentin expression was obvious.Real-time PCR results indicated that the gene expression levels of collagenⅠ andⅢ and TGF-β1 in CFs by exposure to AngⅡ were significantly increased as compared with those in normal group（P〈0.05）.However,AngⅡ-induced collagen Ⅰ and Ⅲ and TGF-β1 mRNA upregulation was inhibited by Res treatment（P〈0.05）.Western blot analysis showed that the protein expressions of collagen Ⅰ and Ⅲ and TGF-β1 in CFs were also increased after exposed to AngⅡ when compared to the normal controls（P〈0.05）.Similarly,AngⅡ-mediated collagenⅠ andⅢand TGF-β1 upregulation was prevented by Res treatment（P〈0.05）.In addition,the phosphorylation level of Smad-3 was enhanced by both interventions（P〈0.05）.However,AngⅡ stimulated TGF-β1 upregulation while Smad-3 phosphorylation was suppressed by Res treatment（P〈0.05）.The secretion levels of collagen Ⅰ and TGF-β1 in CFs supernatant increased significantly in CFs exposed to AngⅡ condition as compared with those in normal condition（P〈0.05）.However,AngⅡ increased collagen Ⅰ and TGF-β1 secretion was prevented by Res intervention（P〈0.05）.Conclusion Resveratrol inhibits the expressions of typeⅠ and typeⅢ collagen induced by AngⅡin cardiac fibroblasts through regulating the TG

关 键 词：心肌纤维化 白藜芦醇 TNF-β/Smads通路 血管紧张素Ⅱ 

分 类 号：R542.2[医药卫生—心血管疾病]