TGF-β induces liver fibrosis via miRNA-181a-mediated down regulation of augmenter of liver regeneration in hepatic stellate cells

• Published in: PloS one Vol. 14; no. 6; p. e0214534
• Main Authors: Gupta, Parul, Sata, Teja Naveen, Yadav, Ajay K, Mishra, Amit, Vats, Nisha, Hossain, Md Musa, Sanal, M G, Venugopal, Senthil Kumar
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 05.06.2019; Public Library of Science (PLoS)
• Subjects: Actin; Actins - metabolism; Apoptosis; Bile; Biology and Life Sciences; Biomarkers - metabolism; Biotechnology; Cell cycle; Cell growth; Cell Line; Collagen; Collagen Type I - metabolism; Down-Regulation; E-cadherin; Fibrosis; Gene Expression Regulation - drug effects; Glyceraldehyde-3-phosphate dehydrogenase; Growth factors; Hepatic Stellate Cells - cytology; Hepatic Stellate Cells - drug effects; Hepatic Stellate Cells - metabolism; Hepatology; Humans; Life sciences; Liver; Liver Cirrhosis - genetics; Liver Cirrhosis - metabolism; Liver transplants; Markers; Medicine and Health Sciences; MicroRNAs; MicroRNAs - genetics; miRNA; Muscles; Overexpression; Oxidoreductases Acting on Sulfur Group Donors - genetics; Oxidoreductases Acting on Sulfur Group Donors - metabolism; Polymerase chain reaction; Proteins; rac1 GTP-Binding Protein - metabolism; Rac1 protein; Reagents; Regeneration; Research and Analysis Methods; Ribonucleic acid; RNA; rRNA 5S; Smooth muscle; Stellate cells; Transfection; Transforming Growth Factor beta - pharmacology; Western blotting; India
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0214534

To study the role of miRNA-181a and augmenter of liver regeneration in TGF-β-induced fibrosis in hepatic stellate cells. 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 actin (α-SMA), rac1, E-cadherin and β-actin. Quantitative RT-PCR was performed for ALR, GAPDH, miRNA-181a or 5S rRNA. TGF-β induced the expression of miRNA-181a, which in turn down-regulated ALR thereby induced the fibrosis markers, such as COLL1A1, α-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 TGFβ-RII and increased expression E-cadherin. TGF-β induced miRNA-181a, which in turn induced fibrosis, at least in part, by inhibiting ALR. ALR inhibited TGF-β action by decreasing the expression of TGFβ-RII, thereby inhibiting miRNA-181a expression and fibrosis markers. ALR could serve as a potential molecule to inhibit liver fibrosis.