miR-200b-containing microvesicles attenuate experimental colitis associated intestinal fibrosis by inhibiting epithelial-mesenchymal transition

J Gastroenterol Hepatol. 2017 Dec;32(12):1966-1974. doi: 10.1111/jgh.13797.

Abstract

Background and aim: Epithelial-mesenchymal transition (EMT), characterized by the decrease of E-cadherin (E-Cad) and increase in vimentin and alpha-smooth muscle actin (α-SMA), was demonstrated to participate in inflammatory bowel disease-related fibrosis. miR-200b plays an anti-fibrosis role in inhibiting EMT by targeting ZEB1 and ZEB2. But the stability of exogenous miR-200b in blood limits its application. Microvesicles (MVs), which can transfer miRNAs among cells and prevent them from degradation, may provide an excellent transport system for the delivery of miR-200b in the treatment of fibrosis.

Methods: Bone marrow mesenchymal stem cells (BMSCs) were transfected with lentivirus to overexpress miR-200b. The MVs packaged with miRNA-200b were harvested for the anti-fibrotic treatment using in vitro (transforming growth factor beta 1-mediated EMT in intestinal epithelial cells: IEC-6) and in vivo (TNBS-induced intestinal fibrosis in rats) models. The pathological morphology was observed, and the fibrosis related proteins, such as E-Cad, vimentin, α-SMA, ZEB1, and ZEB2, were detected.

Results: MiR-200b-MVs would significantly reverse the morphology in TGF-β1-treated IEC-6 cells and improve the TNBS-induced colon fibrosis histologically. The treatment of miR-200b-MVs increased miR-200b levels both in the IEC-6 cells and colon, resulting in a significant prevention EMT and alleviation of fibrosis. The expression of E-Cad was increased, and the expressions of vimentin and α-SMA were decreased. ZBE1 and ZEB2, the targets of miR-200b, were also decreased.

Conclusions: miR-200b could be transferred from genetically modified BMSCs to the target cells or tissue by MVs. The mechanisms of miR-200b-MVs in inhibiting colonic fibrosis were related to suppressing the development of EMT by targeting ZEB1and ZEB2.

Keywords: epithelial-mesenchymal transition; intestinal fibrosis; miR-200b; microvesicles.

MeSH terms

  • Actins / metabolism
  • Animals
  • Cadherins / metabolism
  • Cell-Derived Microparticles*
  • Cells, Cultured
  • Colitis / drug therapy*
  • Colitis / pathology
  • Colitis / physiopathology*
  • Disease Models, Animal
  • Drug Delivery Systems
  • Epithelial-Mesenchymal Transition / drug effects*
  • Epithelial-Mesenchymal Transition / genetics
  • Fibrosis
  • Homeodomain Proteins
  • Intestinal Mucosa / metabolism
  • Intestines / pathology*
  • Male
  • MicroRNAs / administration & dosage*
  • MicroRNAs / metabolism
  • MicroRNAs / pharmacology
  • MicroRNAs / physiology*
  • Molecular Targeted Therapy
  • Rats, Sprague-Dawley
  • Transcription Factors
  • Vimentin / metabolism
  • Zinc Finger E-box-Binding Homeobox 1

Substances

  • Actins
  • Cadherins
  • Homeodomain Proteins
  • MIRN200 microRNA, rat
  • MicroRNAs
  • Transcription Factors
  • Vimentin
  • ZEB1 protein, rat
  • Zinc Finger E-box-Binding Homeobox 1
  • smooth muscle actin, rat