title: Cilostazol improves high glucose-induced impaired angiogenesis in human endothelial progenitor cells and vascular endothelial cells as well as enhances vasculoangiogenesis in hyperglycemic mice mediated by the adenosine monophosphate-activated protein kinase pathway.
abstract: OBJECTIVE
  Cilostazol is an antiplatelet agent with vasodilatory effects that works by increasing intracellular concentrations of cyclic adenosine monophosphate (cAMP). This study investigated the effects of cilostazol in preventing high glucose (HG)-induced impaired angiogenesis and examined the potential mechanisms involving activation of AMP-activated protein kinase (AMPK).
  
  
  METHODS
  Assays for colony formation, adhesion, proliferation, migration, and vascular tube formation were used to determine the effect of cilostazol in HG-treated endothelial progenitor cells (EPCs) or human umbilical vein endothelial cells (HUVECs). Animal-based assays were performed in hyperglycemic ICR mice undergoing hind limb ischemia. An immnunoblotting assay was used to identify the expression and activation of signaling molecules in vitro and in vivo.
  
  
  RESULTS
  Cilostazol treatment significantly restored endothelial function in EPCs and HUVECs through activation of AMPK/acetyl-coenzyme A carboxylase (ACC)-dependent pathways and cAMP/protein kinase A (PKA)-dependent pathways. Recovery of blood flow in the ischemic hind limb and the population of circulating CD34(+) cells were significantly improved in cilostazol-treated mice, and these effects were abolished by local AMPK knockdown. Cilostazol increased the phosphorylation of AMPK/ACC and Akt/endothelial nitric oxide synthase signaling molecules in parallel with or downstream of the cAMP/PKA-dependent signaling pathway in vitro and in vivo.
  
  
  CONCLUSIONS
  Cilostazol prevents HG-induced endothelial dysfunction in EPCs and HUVECs and enhances angiogenesis in hyperglycemic mice by interactions with a broad signaling network, including activation of AMPK/ACC and probably cAMP/PKA pathways.
authors.affiliation: Cardiovascular Research Center, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan; Department of Biological Science, National Sun Yat-Sen University, Kaohsiung, Taiwan.
authors.forename: Shih-Ya
authors.initials: SY
authors.lastname: Tseng
authors.affiliation: Cardiovascular Research Center, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan; Department of Internal Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan. Electronic address: chaoth@mail.ncku.edu.tw.
authors.forename: Ting-Hsing
authors.initials: TH
authors.lastname: Chao
authors.affiliation: Cardiovascular Research Center, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan; Department of Internal Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan.
authors.forename: Yi-Heng
authors.initials: YH
authors.lastname: Li
authors.affiliation: Cardiovascular Research Center, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan; Department of Internal Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan.
authors.forename: Ping-Yen
authors.initials: PY
authors.lastname: Liu
authors.affiliation: Department of Internal Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan.
authors.forename: Cheng-Han
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Department of Biological Science, National Sun Yat-Sen University, Kaohsiung, Taiwan.
authors.forename: Chung-Lung
authors.initials: CL
authors.lastname: Cho
authors.affiliation: Department of Biochemistry and Molecular Biology, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan.
authors.forename: Hua-Lin
authors.initials: HL
authors.lastname: Wu
authors.affiliation: Cardiovascular Research Center, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan; Department of Internal Medicine, National Cheng Kung University College of Medicine and Hospital, Tainan, Taiwan.
authors.forename: Jyh-Hong
authors.initials: JH
authors.lastname: Chen
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D043925:Angiogenesis Inducing Agents / Q000494:pharmacology*; D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D000077407:Cilostazol; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology; D004305:Dose-Response Relationship, Drug; D066026:Endothelial Progenitor Cells / Q000187:drug effects* / Q000201:enzymology / Q000473:pathology; D005947:Glucose / Q000378:metabolism*; D006614:Hindlimb; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007511:Ischemia / Q000188:drug therapy* / Q000201:enzymology / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008813:Mice, Inbred ICR; D018482:Muscle, Skeletal / Q000098:blood supply*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D010766:Phosphorylation; D034622:RNA Interference; D020127:Recovery of Function; D012039:Regional Blood Flow; D015398:Signal Transduction / Q000187:drug effects; D013777:Tetrazoles / Q000494:pharmacology*; D013997:Time Factors; D014162:Transfection
chemical_list: D043925:Angiogenesis Inducing Agents; D013777:Tetrazoles; C548179:Prkaa1 protein, mouse; D055372:AMP-Activated Protein Kinases; C526180:PRKAA1 protein, human; D005947:Glucose; D000077407:Cilostazol