Sphingosine 1-Phosphate Protects Human Umbilical Vein Endothelial Cells from Serum-deprived Apoptosis by Nitric Oxide Production

Young-Guen Kwon, Jeong Ki Min, Ki Mo Kim, Doo Jae Lee, Timothy R. Billiar, Young Myeong Kim

Research output: Contribution to journalArticle

172 Citations (Scopus)

Abstract

Sphingosine 1-phosphate (S1P) can prevent endothelial cell apoptosis. We investigated the molecular mechanisms and signaling pathways by which S1P protects endothelial cells from serum deprivation-induced apoptosis. We show here that human umbilical vein endothelial cells (HUVECs) undergo apoptosis associated with increased DEVDase activity, caspase-3 activation, cytochrome c release, and DNA fragmentation after 24 h of serum deprivation. These apoptotic markers were suppressed by the addition of S1P, the NO donor S-nitroso-N-acetylpenicillamine (100 μM), or caspase-3 inhibitor z-VAD-fmk. The protective effects of S1P were reversed by the nitric-oxide synthase (NOS) inhibitor N-monomethyl-L-arginine, but not by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)oxadiazolo[4,3-α]-quanoxaline-L-one, suggesting that NO, but not cGMP, is responsible for S1P protection from apoptosis. Furthermore, S1P increased NO production by enhancing Ca 2+ -sensitive NOS activity without changes in the eNOS protein level. S1P-mediated cell survival and NO production were suppressed significantly by pretreatment with antisense oligonucleotide of EDG-1 and partially by EDG-3 antisense. S1P-mediated NO production was suppressed by the addition of pertussis toxin, an inhibitor of Gi proteins, the specific inhibitor of phospholipase C (PLC), U73122, and the Ca 2+ chelator BAPTA-AM. These findings indicate that S1P protects HUVECs from apoptosis through the activation of eNOS activity mainly through an EDG-1 and -3/G i /PLC/Ca 2+ signaling pathway.

Original languageEnglish
Pages (from-to)10627-10633
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number14
DOIs
Publication statusPublished - 2001 Apr 6

Fingerprint

Endothelial cells
Human Umbilical Vein Endothelial Cells
Nitric Oxide
Apoptosis
Serum
Type C Phospholipases
Nitric Oxide Synthase
Caspase 3
Endothelial Cells
Chemical activation
S-Nitroso-N-Acetylpenicillamine
sphingosine 1-phosphate
Caspase Inhibitors
Antisense Oligonucleotides
Guanylate Cyclase
Pertussis Toxin
DNA Fragmentation
Chelating Agents
Cytochromes c
Arginine

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kwon, Young-Guen ; Min, Jeong Ki ; Kim, Ki Mo ; Lee, Doo Jae ; Billiar, Timothy R. ; Kim, Young Myeong. / Sphingosine 1-Phosphate Protects Human Umbilical Vein Endothelial Cells from Serum-deprived Apoptosis by Nitric Oxide Production. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 14. pp. 10627-10633.
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abstract = "Sphingosine 1-phosphate (S1P) can prevent endothelial cell apoptosis. We investigated the molecular mechanisms and signaling pathways by which S1P protects endothelial cells from serum deprivation-induced apoptosis. We show here that human umbilical vein endothelial cells (HUVECs) undergo apoptosis associated with increased DEVDase activity, caspase-3 activation, cytochrome c release, and DNA fragmentation after 24 h of serum deprivation. These apoptotic markers were suppressed by the addition of S1P, the NO donor S-nitroso-N-acetylpenicillamine (100 μM), or caspase-3 inhibitor z-VAD-fmk. The protective effects of S1P were reversed by the nitric-oxide synthase (NOS) inhibitor N-monomethyl-L-arginine, but not by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)oxadiazolo[4,3-α]-quanoxaline-L-one, suggesting that NO, but not cGMP, is responsible for S1P protection from apoptosis. Furthermore, S1P increased NO production by enhancing Ca 2+ -sensitive NOS activity without changes in the eNOS protein level. S1P-mediated cell survival and NO production were suppressed significantly by pretreatment with antisense oligonucleotide of EDG-1 and partially by EDG-3 antisense. S1P-mediated NO production was suppressed by the addition of pertussis toxin, an inhibitor of Gi proteins, the specific inhibitor of phospholipase C (PLC), U73122, and the Ca 2+ chelator BAPTA-AM. These findings indicate that S1P protects HUVECs from apoptosis through the activation of eNOS activity mainly through an EDG-1 and -3/G i /PLC/Ca 2+ signaling pathway.",
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Sphingosine 1-Phosphate Protects Human Umbilical Vein Endothelial Cells from Serum-deprived Apoptosis by Nitric Oxide Production. / Kwon, Young-Guen; Min, Jeong Ki; Kim, Ki Mo; Lee, Doo Jae; Billiar, Timothy R.; Kim, Young Myeong.

In: Journal of Biological Chemistry, Vol. 276, No. 14, 06.04.2001, p. 10627-10633.

Research output: Contribution to journalArticle

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AB - Sphingosine 1-phosphate (S1P) can prevent endothelial cell apoptosis. We investigated the molecular mechanisms and signaling pathways by which S1P protects endothelial cells from serum deprivation-induced apoptosis. We show here that human umbilical vein endothelial cells (HUVECs) undergo apoptosis associated with increased DEVDase activity, caspase-3 activation, cytochrome c release, and DNA fragmentation after 24 h of serum deprivation. These apoptotic markers were suppressed by the addition of S1P, the NO donor S-nitroso-N-acetylpenicillamine (100 μM), or caspase-3 inhibitor z-VAD-fmk. The protective effects of S1P were reversed by the nitric-oxide synthase (NOS) inhibitor N-monomethyl-L-arginine, but not by the soluble guanylyl cyclase inhibitor 1H-(1,2,4)oxadiazolo[4,3-α]-quanoxaline-L-one, suggesting that NO, but not cGMP, is responsible for S1P protection from apoptosis. Furthermore, S1P increased NO production by enhancing Ca 2+ -sensitive NOS activity without changes in the eNOS protein level. S1P-mediated cell survival and NO production were suppressed significantly by pretreatment with antisense oligonucleotide of EDG-1 and partially by EDG-3 antisense. S1P-mediated NO production was suppressed by the addition of pertussis toxin, an inhibitor of Gi proteins, the specific inhibitor of phospholipase C (PLC), U73122, and the Ca 2+ chelator BAPTA-AM. These findings indicate that S1P protects HUVECs from apoptosis through the activation of eNOS activity mainly through an EDG-1 and -3/G i /PLC/Ca 2+ signaling pathway.

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