Alcohol induces TGFβ1 via downregulation of miR-1946a in murine lung fibroblast

Sci Rep. 2020 Nov 5;10(1):19089. doi: 10.1038/s41598-020-76148-5.

Abstract

Exaggerated transforming growth factor-beta 1 (TGFβ1) expression worsens fibroproliferation following bleomycin-induced lung injury in alcohol-fed mice. MicroRNA (miR)-1946a is predicted to bind to the TGFβ1 3' untranslated region (UTR), thereby inhibiting its transcription. We hypothesize that alcohol suppresses miR-1946a and induces TGFβ1. Primary murine lung fibroblasts (PLFs) were cultured ± alcohol, miR-1946a mimic or inhibitor, and TGFβ1 signaling inhibitors. miR-1946a was analyzed after alcohol treatment in vitro and in vivo. TGFβ1 expression and TGFβ1 3'UTR-luciferase activity was quantified. We showed that alcohol suppressed miR-1946a in the alcohol-fed mouse lungs and PLFs. MiR-1946a inhibitor increased TGFβ1 expression in the fibroblast. MiR-1946a mimic treatment suppressed TGFβ1 gene expression and TGFβ1 3'UTR activity. Overexpression of miR1946a inhibited alcohol-induced TGFβ1 gene and protein expression as well as alcohol-induced TGFβ1 and α-smooth muscle actin (SMA) protein expression in PLFs. In conclusion, miR-1946a modulates TGFβ1 expression through direct interaction with TGFβ1 3'UTR. These findings identify a novel mechanism by which alcohol induces TGFβ1 in the lung.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 3' Untranslated Regions
  • Actins / genetics
  • Actins / metabolism
  • Alcoholism / genetics
  • Alcoholism / metabolism
  • Alcoholism / pathology
  • Animals
  • Cells, Cultured
  • Down-Regulation / drug effects
  • Ethanol / toxicity*
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • Fibroblasts / pathology
  • Lung / drug effects*
  • Lung / metabolism*
  • Lung / pathology
  • Lung Injury / chemically induced
  • Lung Injury / genetics
  • Lung Injury / metabolism
  • Mice
  • Mice, Inbred C57BL
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism*
  • Transforming Growth Factor beta1 / biosynthesis*
  • Transforming Growth Factor beta1 / genetics*

Substances

  • 3' Untranslated Regions
  • Actins
  • MicroRNAs
  • Tgfb1 protein, mouse
  • Transforming Growth Factor beta1
  • alpha-smooth muscle actin, mouse
  • Ethanol