MicroRNA1 modulates oxLDL-induced hyperlipidemia by down-regulating MLCK and ERK/p38 MAPK pathway

Life Sci. 2014 Jun 27;107(1-2):21-6. doi: 10.1016/j.lfs.2014.04.028. Epub 2014 May 2.

Abstract

Aims: This study was aimed to determine whether microRNA1 (miR1) plays a role in the activation of myosin light chain kinase (MLCK) mediated by oxLDL in human umbilical vein endothelial cells (HUVECs).

Main methods: HUVECs were treated with oxLDL along with a control miR or miR1 mimic. MiR1 expression was assayed by miRNA plate assay kit and mirVana™ miRNA isolation kit. The MLCK protein, transcript, and kinase activity were measured by Western blot, real-time-polymerase chain reaction and γ-(32)P-ATP phosphate incorporation, respectively. In addition, phosphorylation of MLC, ERK and p38 was analyzed by Western blot.

Key findings: The results showed that upon treatment with oxLDL, miR1 expression was decreased, whereas MLCK expression was increased, in a time- and dose-dependent manner. Consistent with this, miR1 mimic prevented MLCK expression and activation and attenuated the phosphorylation of MLC and ERK/p38 in oxLDL-treated HUVECs. Furthermore, we showed that miR1 was able to bind a site located at the 3'un-translational region of MLCK mRNA and inhibited its expression.

Significance: Taken together, this study demonstrated that the effect of miR1 on hyperlipidemia is mediated through down-regulation of MLCK and the ERK/p38 MAPK pathway.

Keywords: HUVECs; Hyperlipidemia; MicroRNA1; Myosin light chain kinase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Blotting, Western
  • Cell Proliferation / drug effects
  • Human Umbilical Vein Endothelial Cells / drug effects*
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Hyperlipidemias / drug therapy
  • Hyperlipidemias / metabolism*
  • Hyperlipidemias / pathology
  • Lipoproteins, LDL / pharmacology*
  • Luciferases / metabolism
  • Membrane Proteins / antagonists & inhibitors
  • Membrane Proteins / metabolism
  • MicroRNAs / physiology*
  • Mitogen-Activated Protein Kinase 1 / metabolism*
  • Mitogen-Activated Protein Kinase 3 / metabolism*
  • Myosin-Light-Chain Kinase / genetics
  • Myosin-Light-Chain Kinase / metabolism*
  • Phosphorylation / drug effects
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction / drug effects
  • p38 Mitogen-Activated Protein Kinases / metabolism*

Substances

  • Lipoproteins, LDL
  • MIRN1 microRNA, human
  • MLC1 protein, human
  • Membrane Proteins
  • MicroRNAs
  • RNA, Messenger
  • oxidized low density lipoprotein
  • Luciferases
  • Myosin-Light-Chain Kinase
  • MAPK1 protein, human
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • p38 Mitogen-Activated Protein Kinases