RESUMO
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 type Ⅰ and type Ⅲ 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 P-Smad-3 were detected by Western blot.The secretion levels of type Ⅰ collagen 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 collagen Ⅰ and Ⅲ 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 collagen Ⅰ and Ⅲand 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 type Ⅰ and type Ⅲ collagen induced by AngⅡ in cardiac fibroblasts through regulating the TGF-β1/Smad3 signaling pathway.