TNF-α elicits phenotypic and functional alterations of vascular smooth muscle cells by miR-155-5p-dependent down-regulation of cGMP-dependent kinase 1

J Biol Chem. 2018 Sep 21;293(38):14812-14822. doi: 10.1074/jbc.RA118.004220. Epub 2018 Aug 13.

Abstract

cGMP-dependent protein kinase 1 (PKG1) plays an important role in nitric oxide (NO)/cGMP-mediated maintenance of vascular smooth muscle cell (VSMC) phenotype and vasorelaxation. Inflammatory cytokines, including tumor necrosis factor-α (TNFα), have long been understood to mediate several inflammatory vascular diseases. However, the underlying mechanism of TNFα-dependent inflammatory vascular disease is unclear. Here, we found that TNFα treatment decreased PKG1 expression in cultured VSMCs, which correlated with NF-κB-dependent biogenesis of miR-155-5p that targeted the 3'-UTR of PKG1 mRNA. TNFα induced VSMC phenotypic switching from a contractile to a synthetic state through the down-regulation of VSMC marker genes, suppression of actin polymerization, alteration of cell morphology, and elevation of cell proliferation and migration. All of these events were blocked by treatment with an inhibitor of miR-155-5p or PKG1, whereas transfection with miR-155-5p mimic or PKG1 siRNA promoted phenotypic modulation, similar to the response to TNFα. In addition, TNFα-induced miR-155-5p inhibited the vasorelaxant response of de-endothelialized mouse aortic vessels to 8-Br-cGMP by suppressing phosphorylation of myosin phosphatase and myosin light chain, both of which are downstream signal modulators of PKG1. Moreover, TNFα-induced VSMC phenotypic alteration and vasodilatory dysfunction were blocked by NF-κB inhibition. These results suggest that TNFα impairs NO/cGMP-mediated maintenance of the VSMC contractile phenotype and vascular relaxation by down-regulating PKG1 through NF-κB-dependent biogenesis of miR-155-5p. Thus, the NF-κB/miR-155-5p/PKG1 axis may be crucial in the pathogenesis of inflammatory vascular diseases, such as atherosclerotic intimal hyperplasia and preeclamptic hypertension.

Keywords: NF-kappa B (NF-κB); cGMP-dependent protein kinase 1; microRNA (miRNA); protein kinase G (PKG); tumor necrosis factor (TNF); vascular smooth muscle cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Actins / metabolism
  • Animals
  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Cyclic GMP-Dependent Protein Kinase Type I / genetics
  • Cyclic GMP-Dependent Protein Kinase Type I / metabolism*
  • Cyclic GMP-Dependent Protein Kinases / metabolism
  • Down-Regulation / physiology*
  • Male
  • Mice, Inbred C57BL
  • MicroRNAs / physiology*
  • Muscle, Smooth, Vascular / cytology*
  • Muscle, Smooth, Vascular / enzymology
  • Muscle, Smooth, Vascular / metabolism
  • NF-kappa B / metabolism
  • Polymerization
  • RNA, Messenger / genetics
  • Tumor Necrosis Factor-alpha / physiology*

Substances

  • 3' Untranslated Regions
  • Actins
  • MicroRNAs
  • Mirn155 microRNA, mouse
  • NF-kappa B
  • RNA, Messenger
  • Tumor Necrosis Factor-alpha
  • Cyclic GMP-Dependent Protein Kinase Type I
  • Cyclic GMP-Dependent Protein Kinases
  • Prkg1 protein, mouse