Ontology highlight
ABSTRACT: Objective
To study the role of miRNA-181a and augmenter of liver regeneration in TGF-?-induced fibrosis in hepatic stellate cells.Methods
LX2 cells were treated with 20 ng/ml TGF-? for 24 h. miRNA-181a, ALR plasmid and empty vectors were transfected using siPORT NeoFx reagent. Cells were harvested after 48 h or 72 h of transfection for protein or RNA analysis. Western blotting was performed for ALR, TGF-? receptor II (TGF?-RII), collagen 1A1 (COLL1A1), alpha-smooth muscle cell actin (?-SMA), rac1, E-cadherin and ?-actin. Quantitative RT-PCR was performed for ALR, GAPDH, miRNA-181a or 5S rRNA.Results
TGF-? induced the expression of miRNA-181a, which in turn down-regulated ALR thereby induced the fibrosis markers, such as COLL1A1, ?-SMA and rac1 in hepatic stellate cells. Over-expression of miRNA-181a down-regulated expression of ALR and up-regulated expression of fibrosis markers. On the other hand, ALR over-expression resulted in a decrease in miRNA-181a expression and fibrosis markers. Over-expression of ALR also inhibited the expression of TGF?-RII and increased expression E-cadherin.Conclusion
TGF-? induced miRNA-181a, which in turn induced fibrosis, at least in part, by inhibiting ALR. ALR inhibited TGF-? action by decreasing the expression of TGF?-RII, thereby inhibiting miRNA-181a expression and fibrosis markers. ALR could serve as a potential molecule to inhibit liver fibrosis.
SUBMITTER: Gupta P
PROVIDER: S-EPMC6550375 | biostudies-literature | 2019
REPOSITORIES: biostudies-literature
Gupta Parul P Sata Teja Naveen TN Yadav Ajay K AK Mishra Amit A Vats Nisha N Hossain Md Musa MM Sanal M G MG Venugopal Senthil Kumar SK
PloS one 20190605 6
<h4>Objective</h4>To study the role of miRNA-181a and augmenter of liver regeneration in TGF-β-induced fibrosis in hepatic stellate cells.<h4>Methods</h4>LX2 cells were treated with 20 ng/ml TGF-β for 24 h. miRNA-181a, ALR plasmid and empty vectors were transfected using siPORT NeoFx reagent. Cells were harvested after 48 h or 72 h of transfection for protein or RNA analysis. Western blotting was performed for ALR, TGF-β receptor II (TGFβ-RII), collagen 1A1 (COLL1A1), alpha-smooth muscle cell ac ...[more]