Cilostazol inhibits oxidative stress-induced premature senescence via upregulation of Sirt1 in human endothelial cells.
OBJECTIVE
Cilostazol, a selective inhibitor of PDE3, has a protective effect on endothelium after ischemic vascular damage, through production of nitric oxide (NO). The purpose of the present study was to clarify the molecular mechanisms underlying the preventive effect of treatment with cilostazol on oxidative stress-induced premature senescence in human endothelial cells.


RESULTS
Prematurely senescent human umbilical vein endothelial cells (HUVECs) were induced by treatment with hydrogen peroxide (H(2)O(2)) as judged by senescence-associated beta-galactosidase assay (SA-betagal), cell morphological appearance, and plasminogen activator inhibitor-1 (PAI-1) expression. Treatment with H(2)O(2) caused 93% of the cells to be SA-betagal positive, whereas 46% of cilostazol (100 micromol/L)-treated cells were positive. HUVECs treated with other cAMP-elevating agents and DETA-NO showed a reduction of SA-betagal-positive cells as well. Cilostazol increased phosphorylation of Akt at Ser(473) and of endothelial nitric oxide synthase (eNOS) at Ser(1177), with a dose-dependent increase in Sirt1 expression. Moreover, the effect of cilostazol on premature senescence was abrogated through inhibition of Sirt1.


CONCLUSIONS
Our results indicated that cilostazol exerted protective effects against endothelial senescence and dysfunction, and enhancement of NO production is a key mediator in upregulation of Sirt1.
Department of Geriatric Medicine, Graduate School of Medicine, University of Tokyo, Japan.
Hidetaka
H
Ota
Masato
M
Eto
Mitsunobu R
MR
Kano
Sumito
S
Ogawa
Katsuya
K
Iijima
Masahiro
M
Akishita
Yasuyoshi
Y
Ouchi
D000107:Acetylation; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D016922:Cellular Senescence / Q000187:drug effects*; D000077407:Cilostazol; D000242:Cyclic AMP / Q000378:metabolism; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3 / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D051379:Mice; D008807:Mice, Inbred BALB C; D009284:Naphthols / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D016877:Oxidants / Q000633:toxicity; D018384:Oxidative Stress; D010269:Paraquat / Q000633:toxicity; D058987:Phosphodiesterase 3 Inhibitors; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010766:Phosphorylation; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D056564:Sirtuin 1; D037761:Sirtuins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013777:Tetrazoles / Q000494:pharmacology*; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015854:Up-Regulation
D001549:Benzamides; D004791:Enzyme Inhibitors; D009284:Naphthols; D016877:Oxidants; D058987:Phosphodiesterase 3 Inhibitors; D010726:Phosphodiesterase Inhibitors; D017395:Plasminogen Activator Inhibitor 1; C051321:SERPINE1 protein, human; C495901:TP53 protein, human; D013777:Tetrazoles; D016159:Tumor Suppressor Protein p53; C439060:sirtinol; D009569:Nitric Oxide; D006861:Hydrogen Peroxide; D000242:Cyclic AMP; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D051057:Proto-Oncogene Proteins c-akt; D054684:Cyclic Nucleotide Phosphodiesterases, Type 3; C447939:SIRT1 protein, human; D056564:Sirtuin 1; D037761:Sirtuins; D000077407:Cilostazol; D010269:Paraquat