Metformin treatment alleviates polycystic ovary syndrome by decreasing the expression of MMP-2 and MMP-9 via H19/miR-29b-3p and AKT/mTOR/autophagy signaling pathways

J Cell Physiol. 2019 Nov;234(11):19964-19976. doi: 10.1002/jcp.28594. Epub 2019 Apr 15.

Abstract

In this study, we aimed to investigate the molecular pathway(s) underlying the effect of metformin (MET) on the expression of matrix metalloproteinase (MMP)-2 and MMP-9. Real-time polymerase chain reaction, Western blot analysis, and gelatin zymography were used to assay the effects of MET on MMP and AMPK signaling pathways. In addition, HTOG cells were treated with miR-29b-3p/a scramble control, H19/a negative control, or MET/PBS to explore possible signaling pathway(s) underlying the inhibitory effect of MET on MMP-2/MMP-9. A rat model of polycystic ovary syndrome (PCOS) was also established to validate the molecular mechanism(s) of MET in vivo. The administration of MET suppressed the expression of MMP-9/MMP-2 and mTOR while increasing the expression of Akt and AMPK, indicating that MET reduced the expression of MMPs via the AMPK signaling pathway. Meanwhile, the H19/miR-29b-3p/MMP-9 and H19/miR-29b-3p/MMP-2 signaling pathways were implicated in PCOS, in which the interactions between H19/miR-29b-3p and MMP-9/MMP-2/miR-29b-3p were confirmed. Furthermore, the administration of MET suppressed the expression of H19 while elevating the expression of miR-29b-3p. And the role of MET in PCOS was also confirmed in vivo via examining the activity of H19 and AMPK signaling pathways in cell or serum samples collected from PCOS rats. MET exhibits a therapeutic effect in the treatment of PCOS by reducing the expression of MMPs.

Keywords: Akt; H19; MET; MMP-2; MMP-9; PCOS; autophagy; mTOR; miR-29b-3p.

MeSH terms

  • Adenylate Kinase / metabolism
  • Animals
  • Autophagy* / drug effects
  • Base Sequence
  • Cell Line
  • Down-Regulation / drug effects
  • Down-Regulation / genetics
  • Female
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism*
  • Matrix Metalloproteinase 9 / genetics
  • Matrix Metalloproteinase 9 / metabolism*
  • Metformin / pharmacology
  • Metformin / therapeutic use*
  • Polycystic Ovary Syndrome / blood
  • Polycystic Ovary Syndrome / drug therapy*
  • Polycystic Ovary Syndrome / enzymology*
  • Proto-Oncogene Proteins c-akt / metabolism
  • Proto-Oncogene Proteins c-met / metabolism
  • RNA, Long Noncoding / genetics
  • RNA, Long Noncoding / metabolism*
  • Rats, Sprague-Dawley
  • Signal Transduction* / drug effects
  • TOR Serine-Threonine Kinases / metabolism

Substances

  • H19 long non-coding RNA
  • RNA, Long Noncoding
  • Metformin
  • Proto-Oncogene Proteins c-met
  • Proto-Oncogene Proteins c-akt
  • TOR Serine-Threonine Kinases
  • Adenylate Kinase
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9