title: Down-regulation of miR-34a alleviates mesangial proliferation in vitro and glomerular hypertrophy in early diabetic nephropathy mice by targeting GAS1.
abstract: OBJECTIVE
  Diabetic nephropathy (DN) is a major diabetic complication characterized by mesangial proliferation and glomerular hypertrophy. MicroRNAs might play an important role in these pathological processes. The aim of this study is to examine the possible association of miR-34a as one of the microRNAs with DN and underlying mechanisms in vitro and in vivo.
  
  
  METHODS
  According to previous results of microarray which compared the different microRNAs between diabetic and normal control mice, miR-34a was chosen and its expression was detected by qRT-PCR. Cell viability was then assessed using Cell Counting Kit-8 (CCK8) and 5-ethynyl-20-deoxyuridine (EDU) incorporation. Antagomir was injected in db/db mice to down regulate miR-34a. Average diameter of glomeruli was analyzed by periodic acid-Schiff (PAS) stain of kidney. Luciferase gene report assay was then performed to identify the target gene of miR-34a. Additional immunoblotting and immunohistochemical analyses were implemented to verify the expression level of growth arrest-specific 1 (GAS1).
  
  
  RESULTS
  MiR-34a expression level was increased under high glucose condition in vitro and in vivo. Down-regulation of miR-34a inhibits mice mesangial cells (MMCs) proliferation in vitro and alleviates glomerular hypertrophy in vivo. GAS1 was proved to be the target of miR-34a through luciferase report. Moreover, up-regulation of GAS1 expression was observed in the presence of miR-34a antagomir as compared with miR-34a antagomir-NC in high-glucose-treated MMCs and db/db mice, respectively.
  
  
  CONCLUSIONS
  MiR-34a regulated mesangial proliferation and glomerular hypertrophy by directly inhibiting GAS1 in early DN.
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Le
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Department of Cardiovascular Surgery, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Siyi
authors.initials: S
authors.lastname: He
authors.affiliation: Division of Molecular Cardiology, Cardiovascular Research Institute, College of Medicine, Texas A&M Health Science Center, Central Texas Veterans Health Care System, Temple, TX 76504, USA.
authors.forename: Shaodong
authors.initials: S
authors.lastname: Guo
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Xie
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Xin
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Yu
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Qiu
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China.
authors.forename: Di
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Clinical Drug Trials Agency, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. Electronic address: zhouswxinqiao@163.com.
authors.forename: Shiwen
authors.initials: S
authors.lastname: Zhou
authors.affiliation: Center of Medical Experiment & Technology, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. Electronic address: zhangkebin12@163.com.
authors.forename: Kebin
authors.initials: K
authors.lastname: Zhang
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D049109:Cell Proliferation / Q000502:physiology*; D002470:Cell Survival; D002478:Cells, Cultured; D003924:Diabetes Mellitus, Type 2 / Q000150:complications* / Q000378:metabolism; D003928:Diabetic Nephropathies / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004195:Disease Models, Animal; D015536:Down-Regulation / Q000502:physiology*; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000494:pharmacology; D006984:Hypertrophy / Q000517:prevention & control; D066298:In Vitro Techniques; D007678:Kidney Glomerulus / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D008297:Male; D050527:Mesangial Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*
chemical_list: D018797:Cell Cycle Proteins; D058851:GPI-Linked Proteins; C501180:Gas1 protein, mouse; C551848:MIRN34a microRNA, mouse; D035683:MicroRNAs; D005947:Glucose
keywords: Diabetic nephropathy; GAS1; Glomerular hypertrophy; miR-34a