Tauroursodeoxycholic acid through modulating endoplasmic reticulum stress on inhibition of transforming growth factor β1-induced fibrosis in cardiac fibroblasts
Tauroursodeoxycholic acid through modulating endoplasmic reticulum stress on inhibition of transforming growth factor β1-induced fibrosis in cardiac fibroblasts
Objective To investigate the effect of taurodeoxycholic acid(TUDCA)on the activation of cardiac fibroblasts(CFs)induced by transforming growth factor β1(TGF-β1)and its related mechanisms,and to identify a new therapeutic target for TUDCA in the treatment of diabetic myocardial fibrosis.Methods The CFs and cardiomyocytes of primary neonatal SD rats were extracted,and the purity of CFs was identified by immunofluorescence.CFs cultured with high glucose were treated with TGF-β1 for different duration.The expression of endoplasmic reticulum stress(ERS)pathway related proteins and fibrosis markers were detected by Western blot to explore the effect of TGF-β1 on CFs activation and ERS related pathway.Western blot was used to detect the expression of homocysteine-inducible,ERS-inducible,ubiquitin-like domain member 1(Herpud1)in CFs and cardiomyocytes.The role of Herpud1 in TGF-β1-induced CFs activation was explored by silencing the expression of Herpud1 and detecting the expression of CFs fibrosis markers after silencing Herpud1 by Transwell and Western blot.CCK-8 was used to detect the activity of CFs treated with different concentrations of TUDCA.Immunofluorescence and Western blot were used to detect the effects of TUDCA on the expression of Herpud1,glucose regulated protein 78(GRP78),transcriptional activator 6(ATF6),α-smooth muscle actin(α-SMA),vimentin(Vimentin)and type Ⅰ collagen(Collagen Ⅰ)in CFs induced by TGF-β1.In order to further clarify the role of Herpud1 in the inhibition of CFs activation by TUDCA,the expression of related proteins after reaching Herpud1 was detected by Western blot.Results Based on the successful construction of a fibrosis model of CFs,TGF-β1 was proved to induce CFs activation as well as the expression of ERS pathway-related proteins.Expression of Herpud1 was higher in CFs than in cardiomyocytes.Knockdown of Herpud1 showed inhibition of CFs activation induced by TGF-β1.TUDCA significantly reduced TGF-β1-induced activation of CFs in GRP78,ATF6,α-SMA,Vimentin and Collagen Ⅰ expression levels.In addition,overexpression of Herpud1 reversed the inhibition of TGF-β1-induced activation of CFs by TUDCA.Conclusions Down-regulation of Herpud1 gene expression is one of the mechanisms by which TUDCA inhibits TGF-β1-induced cellular fibrosis in CFs.
维普
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