Effects and mechanism of UBC9 in liver fibrosis
10.3969/j.issn.1006-5725.2017.13.003
- VernacularTitle:类泛素缀和酶9在肝纤维化中的作用及机制
- Author:
Sufen FANG
;
Wuhua GUO
- Keywords:
liver fibrosis;
UBC9;
hepatic stellate cells;
TGF-β1/Smad pathway
- From:
The Journal of Practical Medicine
2017;33(13):2079-2082
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the roles of ubiquitin-conjugating enzyme(UBC9)in HSCs activa-tion and liver fibrosis. Methods Western blot was used to analyze the expression of UBC9 under the stimulus of different concentrations of TGF-β1. The effective shRNA-targeting UBC9 gene was synthesized and HCSs were in-stantaneously transfected using lipofectamine method. Non-specific shRNA-transfected group cells and shRNA-tar-geting UBC9-transfected group cells were set up. The mRNA and protein levels of UBC9 were determined with Quantitative Real-Time PCR and Western blot. Western blot also used to examine the expression level of collagenⅠ,α-SMA and P-smad3 after transfection of UBC9 shRNA into HCSs and CCK-8 assay was used to detect cell proliferative capacity after transfection. Results UBC9 expression was significantly up-regulated in TGF-β1-treat-ed HSCs. Knockdown of UBC9 significantly inhibited TGF-β1-induced HSCs proliferation,as well as decreased the expression levels of a-SMA and collagen I. Furthermore ,knockdown of UBC9 attenuated the phosphorylation of Smad3 in the presence of TGF-β1. Conclusions UBC9 may function as a novel regulator to modulate HSC activa-tion,potentially by inhibiting the TGF-β1/Smad3 signaling pathway,which reveals novel mechanistic insights into the anti-fibrotic effect of UBC9.
