Resveratrol inhibits pulmonary fibrosis by regulating miR-21 through MAPK/AP-1 pathways

Biomed Pharmacother. 2018 Sep:105:37-44. doi: 10.1016/j.biopha.2018.05.104. Epub 2018 May 26.

Abstract

Objective: To explore the molecular mechanism of Res in regulation of pulmonary fibrosis (PF).

Methods: Rats were injected with bleomycin (BLM) to establish a PF model and treated with resveratrol (Res) and/or miR-21 agomir. After 14 days, lung tissues were collected for Hematoxylin-eosin and Masson's staining, and real-time quantitative polymerase chain reaction and Western blot were performed to detect fibrosis-related protein expression and the activation of the TGF-β1/Smad pathway. In vitro, MRC-5 cells were pretreated with TGF-β1, Res, and/or miR-21 agomir. After 48 h, total soluble collagen was detected with a Sircol Soluble Collagen Assay. Subsequently, a miR-21 mimic was transfected into MRC-5 cells, and a luciferase reporter assay was employed to verify whether miR-21 targeted Smad7.

Results: Res reversed the increased levels of miR-21 induced by BLM and alleviated serious PF symptoms, but agomiR-21 treatment effectively impaired the above manifestations. In vivo, miR-21 inhibited the decreases of TGF-β1 and p-Smad2/3 that were induced by Res. In vitro, miR-21 significantly disrupted the positive effect of Res on TGF-β-induced collagen deposition, as well as the levels of Fn, α-SMA, p-Smad2, and Smad7. In addition, Smad7 was found to be a direct target of miR-21-5p. TGF-β stimulation led to an enormous increase in p-c-Jun, c-Jun, and c-Fos, which were significantly reduced by Res. Finally, miR-21 sharply reduced the increased phosphorylation levels of ERK, JNK and p38 that were induced by Res.

Conclusion: Res inhibits BLM-induced PF by regulating miR-21 through MAPK/AP-1 pathways.

Keywords: MAPK/AP-1 pathway; Resveratrol; TGF-β1/Smad; miR-21; pulmonary fibrosis.

MeSH terms

  • Animals
  • Antioxidants / administration & dosage
  • Antioxidants / therapeutic use*
  • Disease Models, Animal
  • Female
  • Gene Expression Regulation / drug effects
  • MAP Kinase Signaling System / drug effects*
  • Male
  • MicroRNAs / genetics*
  • Pulmonary Fibrosis / drug therapy*
  • Pulmonary Fibrosis / metabolism
  • Pulmonary Fibrosis / pathology
  • Rats, Sprague-Dawley
  • Resveratrol
  • Stilbenes / administration & dosage
  • Stilbenes / therapeutic use*
  • Transcription Factor AP-1 / metabolism*

Substances

  • Antioxidants
  • MIRN21 microRNA, human
  • MicroRNAs
  • Stilbenes
  • Transcription Factor AP-1
  • Resveratrol