title: MicroRNA1 modulates oxLDL-induced hyperlipidemia by down-regulating MLCK and ERK/p38 MAPK pathway.
abstract: OBJECTIVE
  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).
  
  
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China; Key Laboratory of Gene Research of Anhui Province, Hefei, Anhui, People's Republic of China. Electronic address: aydzhq@126.com.
authors.forename: Hua-Qing
authors.initials: HQ
authors.lastname: Zhu
authors.affiliation: Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Pharmacology, Anhui Medical University, Hefei, Anhui, People's Republic of China.
authors.forename: Liu-Yi
authors.initials: LY
authors.lastname: Dong
authors.affiliation: Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China; Key Laboratory of Gene Research of Anhui Province, Hefei, Anhui, People's Republic of China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Laboratory of Molecular Biology and Department of Biochemistry, Anhui Medical University, Hefei, Anhui, People's Republic of China; Key Laboratory of Gene Research of Anhui Province, Hefei, Anhui, People's Republic of China. Electronic address: wangyuan@ahmu.edu.cn.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006949:Hyperlipidemias / Q000188:drug therapy / Q000378:metabolism* / Q000473:pathology; D008077:Lipoproteins, LDL / Q000494:pharmacology*; D008156:Luciferases / Q000378:metabolism; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D035683:MicroRNAs / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D008077:Lipoproteins, LDL; C544547:MIRN1 microRNA, human; C425069:MLC1 protein, human; D008565:Membrane Proteins; D035683:MicroRNAs; D012333:RNA, Messenger; C099252:oxidized low density lipoprotein; D008156:Luciferases; D009219:Myosin-Light-Chain Kinase; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D048051:p38 Mitogen-Activated Protein Kinases
keywords: HUVECs; Hyperlipidemia; MicroRNA1; Myosin light chain kinase