Obesity regulates miR-467/HoxA10 axis on osteogenic differentiation and fracture healing by BMSC-derived exosome LncRNA H19

J Cell Mol Med. 2021 Feb;25(3):1712-1724. doi: 10.1111/jcmm.16273. Epub 2021 Jan 20.

Abstract

This study explored the therapeutic effect of bone marrow mesenchymal stem cell-derived exosomes on the treatment of obesity-induced fracture healing. Quantitative real-time PCR was used to detect the expression of lncRNA H19, miR-467 and Hoxa10 and combined with WB detection to detect osteogenic markers (RUNX2, OPN, OCN). Determine whether exosomes have entered BMSCs by immunofluorescence staining. Alkaline phosphatase (ALP) and alizarin red staining (ARS) staining were used to detect ALP activity and calcium deposition. We found that high-fat treatment can inhibit the secretion of BMSCs-derived exosomes and affect the expression of H19 carried by them. In vivo and in vitro experiments show that high-fat or obesity factors can inhibit the expression of osteogenic markers and reduce the staining activity of ALP and ARS. The treatment of exosomes from normal sources can reverse the phenomenon of osteogenic differentiation and abnormal fracture healing. Further bioinformatics analysis found that miR-467 as a regulatory molecule of lncRNA H19 and Hoxa10, and we verified the targeting relationship of the three through dual luciferase report experiments. Further, we found similar phenomena in ALP and ARS staining. Bone marrow mesenchymal stem cell-derived exosomes improve fracture healing caused by obesity.

Keywords: bone marrow mesenchymal stem cells; exosomes; fracture healing; obesity.

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Disease Models, Animal
  • Exosomes / metabolism*
  • Fracture Healing / genetics*
  • Homeobox A10 Proteins / genetics*
  • Humans
  • Male
  • Mesenchymal Stem Cells / metabolism*
  • Mice
  • MicroRNAs / genetics*
  • Obesity / genetics
  • Obesity / metabolism
  • Osteogenesis / genetics*
  • RNA, Long Noncoding / genetics*

Substances

  • H19 long non-coding RNA
  • Homeobox A10 Proteins
  • MicroRNAs
  • RNA, Long Noncoding
  • HOXA10 protein, human