Quercetin regulates fibrogenic responses of endometrial stromal cell by upregulating miR-145 and inhibiting the TGF-β1/Smad2/Smad3 pathway

Acta Histochem. 2020 Oct;122(7):151600. doi: 10.1016/j.acthis.2020.151600. Epub 2020 Aug 29.

Abstract

Objectives: Aim of this study is to explore whether quercetin can inhibit the enlarged fibrogenic responses of endometrial stromal cells by increasing the level of microRNA-145 (miR-145) and mediating the TGFβ1/Smad2/Smad3 signaling pathway, and to discuss the mechanism of signal transduction, further to provide experimental basis for revealing the pathophysiological mechanism and seeking new strategies for effective prevention and treatment of endometrial fibrosis.

Methods: The expression levels of miR-145 and TGF-β receptor 2 (TGFBR2) were detected by RT-qPCR analysis. Expressions of α-smooth muscle actin (α-SMA) and vimentin were examined by immunofluorescence staining. Cell viability was measured by MTT assay. The protein expression of collagen type 1 alpha 1 (Col1a1), α-SMA, fibronectin (FN), TGFBR2, transforming growth factor (TGF-β1), Smad2/3, phospho-Smad2/3 (p-Smad2/3) were detected by western blot analysis. The interaction between miR-145 and TGFBR2 was confirmed by dual-luciferase reporter gene assay.

Results: The expression level of miR-145 was decreased, whereas TGFBR2 was increased in intrauterine adhesion tissue. The expression levels of COL1A1, α-SMA, FN, TGFBR2, and p-Smad2/3 were increased, whereas miR-145 and cell proliferation were decreased in human endometrial stromal cells (hESCs) in response to TGF-β1 stimulation in a time and dose-dependent manner, which could be reversed by quercetin. Furthermore, quercetin regulates cell fibrogenic responses of endometrial stromal cells via miR-145/TGF-β1/Smad2/Smad3 pathway.

Conclusions: These findings indicated that quercetin have a significant anti-fibrotic effect and could upregulate miR-145 and inhibit activation of TGF-β1/Smad2/Smad3 pathway to regulate TGF-β1 induced fibrogenic responses of endometrial stromal cells, which may serve as a potential therapeutic agent for endometrial fibrosis.

Keywords: Endometrial fibrosis; TGF-β1; TGFBR2; microRNA-145; quercetin.

MeSH terms

  • Adult
  • Female
  • Humans
  • Male
  • MicroRNAs / drug effects*
  • Quercetin / pharmacology*
  • Receptor, Transforming Growth Factor-beta Type II / drug effects*
  • Smad2 Protein / drug effects*
  • Smad3 Protein / drug effects*
  • Stromal Cells / drug effects
  • Transforming Growth Factor beta1 / drug effects
  • Transforming Growth Factor beta1 / genetics

Substances

  • MIRN145 microRNA, human
  • MicroRNAs
  • SMAD2 protein, human
  • SMAD3 protein, human
  • Smad2 Protein
  • Smad3 Protein
  • TGFB1 protein, human
  • Transforming Growth Factor beta1
  • Quercetin
  • Receptor, Transforming Growth Factor-beta Type II
  • TGFBR2 protein, human