title: Cellular repressor of E1A-stimulated gene overexpression in bone mesenchymal stem cells protects against rat myocardial infarction.
abstract: BACKGROUND
  Bone mesenchymal stem cell (BMSC) therapy has modest success in ischemic heart disease but has been limited by poor survival in diseased microenvironments. Cellular repressor of E1A-stimulated genes (CREG) can prevent BMSCs from apoptosis in vitro; however, the effects of CREG-modified BMSCs on ischemic heart disease and the related mechanism remain undefined. Therefore, we designed to study the cardioprotective effects of CREG overexpression in BMSCs ((CREG)BMSCs) after transplantation into infarcted heart of rats.
  
  
  METHODS
  In vivo studies, 50 μl PBS or 1.5×10(6)(Norm)BMSCs, (GFP)BMSCs or (CREG)BMSCs were implanted intramyocardially in myocardial infarction rat models. 3 or 14 days later, cardiac function, fibrosis, apoptosis and angiogenesis were analyzed by echocardiography, masson, western blot and immunofluorescence staining, respectively. ELISA, western blot and matrigel assay were used in vitro to detect vascular endothelial growth factor (VEGF) secretion, signaling molecule expression, and angiogenic tube formation.
  
  
  RESULTS
  In vivo, prolonged cardiac function (14d LVEF: 50.87 ± 0.94%; LVFS: 23.41 ± 1.12%), decreased fibrosis (14d Fibrotic area: 27.37 ± 1.03%) and apoptosis and increased angiogenesis were observed in (CREG)BMSCs, compared with other groups. In vivo and in vitro, VEGF secretion from (CREG)BMSCs was markedly enhanced. In vitro, angiogenic tube formation in (CREG)BMSC supernatants significantly increased. Moreover, CREG activated hypoxia-inducible factor-1α (HIF-1α), but not HIF-1β. Knockdown of HIF-1α with siRNA decreased VEGF secretion and angiogenic tube formation. Notably, CREG did not influence HIF-1α mRNA synthesis but inhibited the expression of Von Hippel-Lindau (VHL), a key protein that regulates HIF-1α degradation.
  
  
  CONCLUSIONS
  The (CREG)BMSC transplantation, directly or indirectly, may promote VEGF's anti-apoptosis and angiogenesis via the inhibition of VHL-mediated HIF-1α degradation, consequently protecting against myocardial infarction.
authors: Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China; Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Chengfei
authors: C
authors: Peng
authors: Department of Cardiology, Chengdu Military General Hospital, Chengdu 610083, China.
authors: Haifeng
authors: H
authors: Pei
authors: Department of Interventional Radiology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors: Feipeng
authors: F
authors: Wei
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Xiaoxiang
authors: X
authors: Tian
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Jie
authors: J
authors: Deng
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Chenghui
authors: C
authors: Yan
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Yang
authors: Y
authors: Li
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Mingyu
authors: M
authors: Sun
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China.
authors: Jian
authors: J
authors: Zhang
authors: Department of Cardiology, Graduate School, Third Military Medical University, Chongqing 400038, China.
authors: Dan
authors: D
authors: Liu
authors: Department of Cardiology, Graduate School, Third Military Medical University, Chongqing 400038, China.
authors: Jingjing
authors: J
authors: Rong
authors: Department of Cardiology, Tangdu Hospital, Fourth Military Medical University, Xi'an 710038, China.
authors: Jie
authors: J
authors: Wang
authors: Center for Translational Medicine, Temple University School of Medicine, Philadelphia 19104, USA.
authors: Erhe
authors: E
authors: Gao
authors: Department of Surgery, Rutgers University Robert Wood Johnson Medical School, New Brunswick 08904, USA.
authors: Shaohua
authors: S
authors: Li
authors: Cardiovascular Research Institute, Department of Cardiology, General Hospital of Shenyang Military Region, Shenyang 110016, China. Electronic address: hanyaling2014@126.com.
authors: Yaling
authors: Y
authors: Han
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D001842:Bone and Bones / Q000166:cytology*; D015870:Gene Expression; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D023421:Models, Animal; D009203:Myocardial Infarction / Q000628:therapy*; D017202:Myocardial Ischemia / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000166:cytology; D018919:Neovascularization, Physiologic; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000378:metabolism
chemical_list: C495367:Creg1 protein, rat; C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D012097:Repressor Proteins; D042461:Vascular Endothelial Growth Factor A; D051794:Von Hippel-Lindau Tumor Suppressor Protein
keywords: BMSCs; CREG; HIF-1α; Myocardial infarction; VHL