Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53-dependent Noxa expression through the formation of S-nitrosohomocysteine

Seon Jin Lee, Ki Mo Kim, Seung Namkoong, Chun Ki Kim, Yun Chul Kang, Hansoo Lee, Kwon Soo Ha, Jeong A. Han, Hun Taeg Chung, Young-Guen Kwon, Young Myeong Kim

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Hyperhomocysteinemia is believed to induce endothelial dysfunction and promote atherosclerosis; however, the pathogenic mechanism has not been clearly elucidated. In this study, we examined the molecular mechanism by which homocysteine (HCy) causes endothelial cell apoptosis and by which nitric oxide (NO) affects HCy-induced apoptosis. Our data demonstrated that HCy caused caspase-dependent apoptosis in cultured human umbilical vein endothelial cells, as determined by cell viability, nuclear condensation, and caspase-3 activation and activity. These apoptotic characteristics were correlated with reactive oxygen species (ROS) production, lipid peroxidation, p53 and Noxa expression, and mitochondrial cytochrome c release following HCy treatment. HCy also induced p53 and Noxa expression and apoptosis in endothelial cells from wild type mice but not in the p53-deficient cells. The NO donor S-nitroso-N- acetylpenicillamine, adenoviral transfer of inducible NO synthase gene, and antioxidants (α-tocopherol and superoxide dismutase plus catalase) but not oxidized SNAP, 8-Br-cGMP, nitrite, and nitrate, suppressed ROS prodection, p53-dependent Noxa expression, and apoptosis induced by HCy. The cytotoxic effect of HCy was decreased by small interfering RNA-mediated suppression of Noxa expression, indicating that Noxa upregulation plays an important role in HCy-induced endothelial cell apoptosis. Overexpression of inducible NO synthase increased the formation of S-nitroso-HCy, which was inhibited by the NO synthase inhibitor N-monomethyl-L-arginine. Moreover, S-nitroso-HCy did not increase ROS generation, p53-dependent Noxa expression, and apoptosis. These results suggest that upregulation of p53-dependent Noxa expression may play an important role in the pathogenesis of atherosclerosis induced by HCy and that an increase in vascular NO production may prevent HCy-induced endothelial dysfunction by S-nitrosylation.

Original languageEnglish
Pages (from-to)5781-5788
Number of pages8
JournalJournal of Biological Chemistry
Volume280
Issue number7
DOIs
Publication statusPublished - 2005 Feb 18

Fingerprint

Noxae
Endothelial cells
Homocysteine
Nitric Oxide
Down-Regulation
Endothelial Cells
Apoptosis
Reactive Oxygen Species
Nitric Oxide Synthase Type II
S-nitrosohomocysteine
Atherosclerosis
Up-Regulation
S-Nitroso-N-Acetylpenicillamine
Hyperhomocysteinemia
Tocopherols
Nitric Oxide Donors
Human Umbilical Vein Endothelial Cells
Caspases
Nitrites
Cytochromes c

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lee, Seon Jin ; Kim, Ki Mo ; Namkoong, Seung ; Kim, Chun Ki ; Kang, Yun Chul ; Lee, Hansoo ; Ha, Kwon Soo ; Han, Jeong A. ; Chung, Hun Taeg ; Kwon, Young-Guen ; Kim, Young Myeong. / Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53-dependent Noxa expression through the formation of S-nitrosohomocysteine. In: Journal of Biological Chemistry. 2005 ; Vol. 280, No. 7. pp. 5781-5788.
@article{587ccd98460c4860ae9b44674215926e,
title = "Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53-dependent Noxa expression through the formation of S-nitrosohomocysteine",
abstract = "Hyperhomocysteinemia is believed to induce endothelial dysfunction and promote atherosclerosis; however, the pathogenic mechanism has not been clearly elucidated. In this study, we examined the molecular mechanism by which homocysteine (HCy) causes endothelial cell apoptosis and by which nitric oxide (NO) affects HCy-induced apoptosis. Our data demonstrated that HCy caused caspase-dependent apoptosis in cultured human umbilical vein endothelial cells, as determined by cell viability, nuclear condensation, and caspase-3 activation and activity. These apoptotic characteristics were correlated with reactive oxygen species (ROS) production, lipid peroxidation, p53 and Noxa expression, and mitochondrial cytochrome c release following HCy treatment. HCy also induced p53 and Noxa expression and apoptosis in endothelial cells from wild type mice but not in the p53-deficient cells. The NO donor S-nitroso-N- acetylpenicillamine, adenoviral transfer of inducible NO synthase gene, and antioxidants (α-tocopherol and superoxide dismutase plus catalase) but not oxidized SNAP, 8-Br-cGMP, nitrite, and nitrate, suppressed ROS prodection, p53-dependent Noxa expression, and apoptosis induced by HCy. The cytotoxic effect of HCy was decreased by small interfering RNA-mediated suppression of Noxa expression, indicating that Noxa upregulation plays an important role in HCy-induced endothelial cell apoptosis. Overexpression of inducible NO synthase increased the formation of S-nitroso-HCy, which was inhibited by the NO synthase inhibitor N-monomethyl-L-arginine. Moreover, S-nitroso-HCy did not increase ROS generation, p53-dependent Noxa expression, and apoptosis. These results suggest that upregulation of p53-dependent Noxa expression may play an important role in the pathogenesis of atherosclerosis induced by HCy and that an increase in vascular NO production may prevent HCy-induced endothelial dysfunction by S-nitrosylation.",
author = "Lee, {Seon Jin} and Kim, {Ki Mo} and Seung Namkoong and Kim, {Chun Ki} and Kang, {Yun Chul} and Hansoo Lee and Ha, {Kwon Soo} and Han, {Jeong A.} and Chung, {Hun Taeg} and Young-Guen Kwon and Kim, {Young Myeong}",
year = "2005",
month = "2",
day = "18",
doi = "10.1074/jbc.M411224200",
language = "English",
volume = "280",
pages = "5781--5788",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "7",

}

Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53-dependent Noxa expression through the formation of S-nitrosohomocysteine. / Lee, Seon Jin; Kim, Ki Mo; Namkoong, Seung; Kim, Chun Ki; Kang, Yun Chul; Lee, Hansoo; Ha, Kwon Soo; Han, Jeong A.; Chung, Hun Taeg; Kwon, Young-Guen; Kim, Young Myeong.

In: Journal of Biological Chemistry, Vol. 280, No. 7, 18.02.2005, p. 5781-5788.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Nitric oxide inhibition of homocysteine-induced human endothelial cell apoptosis by down-regulation of p53-dependent Noxa expression through the formation of S-nitrosohomocysteine

AU - Lee, Seon Jin

AU - Kim, Ki Mo

AU - Namkoong, Seung

AU - Kim, Chun Ki

AU - Kang, Yun Chul

AU - Lee, Hansoo

AU - Ha, Kwon Soo

AU - Han, Jeong A.

AU - Chung, Hun Taeg

AU - Kwon, Young-Guen

AU - Kim, Young Myeong

PY - 2005/2/18

Y1 - 2005/2/18

N2 - Hyperhomocysteinemia is believed to induce endothelial dysfunction and promote atherosclerosis; however, the pathogenic mechanism has not been clearly elucidated. In this study, we examined the molecular mechanism by which homocysteine (HCy) causes endothelial cell apoptosis and by which nitric oxide (NO) affects HCy-induced apoptosis. Our data demonstrated that HCy caused caspase-dependent apoptosis in cultured human umbilical vein endothelial cells, as determined by cell viability, nuclear condensation, and caspase-3 activation and activity. These apoptotic characteristics were correlated with reactive oxygen species (ROS) production, lipid peroxidation, p53 and Noxa expression, and mitochondrial cytochrome c release following HCy treatment. HCy also induced p53 and Noxa expression and apoptosis in endothelial cells from wild type mice but not in the p53-deficient cells. The NO donor S-nitroso-N- acetylpenicillamine, adenoviral transfer of inducible NO synthase gene, and antioxidants (α-tocopherol and superoxide dismutase plus catalase) but not oxidized SNAP, 8-Br-cGMP, nitrite, and nitrate, suppressed ROS prodection, p53-dependent Noxa expression, and apoptosis induced by HCy. The cytotoxic effect of HCy was decreased by small interfering RNA-mediated suppression of Noxa expression, indicating that Noxa upregulation plays an important role in HCy-induced endothelial cell apoptosis. Overexpression of inducible NO synthase increased the formation of S-nitroso-HCy, which was inhibited by the NO synthase inhibitor N-monomethyl-L-arginine. Moreover, S-nitroso-HCy did not increase ROS generation, p53-dependent Noxa expression, and apoptosis. These results suggest that upregulation of p53-dependent Noxa expression may play an important role in the pathogenesis of atherosclerosis induced by HCy and that an increase in vascular NO production may prevent HCy-induced endothelial dysfunction by S-nitrosylation.

AB - Hyperhomocysteinemia is believed to induce endothelial dysfunction and promote atherosclerosis; however, the pathogenic mechanism has not been clearly elucidated. In this study, we examined the molecular mechanism by which homocysteine (HCy) causes endothelial cell apoptosis and by which nitric oxide (NO) affects HCy-induced apoptosis. Our data demonstrated that HCy caused caspase-dependent apoptosis in cultured human umbilical vein endothelial cells, as determined by cell viability, nuclear condensation, and caspase-3 activation and activity. These apoptotic characteristics were correlated with reactive oxygen species (ROS) production, lipid peroxidation, p53 and Noxa expression, and mitochondrial cytochrome c release following HCy treatment. HCy also induced p53 and Noxa expression and apoptosis in endothelial cells from wild type mice but not in the p53-deficient cells. The NO donor S-nitroso-N- acetylpenicillamine, adenoviral transfer of inducible NO synthase gene, and antioxidants (α-tocopherol and superoxide dismutase plus catalase) but not oxidized SNAP, 8-Br-cGMP, nitrite, and nitrate, suppressed ROS prodection, p53-dependent Noxa expression, and apoptosis induced by HCy. The cytotoxic effect of HCy was decreased by small interfering RNA-mediated suppression of Noxa expression, indicating that Noxa upregulation plays an important role in HCy-induced endothelial cell apoptosis. Overexpression of inducible NO synthase increased the formation of S-nitroso-HCy, which was inhibited by the NO synthase inhibitor N-monomethyl-L-arginine. Moreover, S-nitroso-HCy did not increase ROS generation, p53-dependent Noxa expression, and apoptosis. These results suggest that upregulation of p53-dependent Noxa expression may play an important role in the pathogenesis of atherosclerosis induced by HCy and that an increase in vascular NO production may prevent HCy-induced endothelial dysfunction by S-nitrosylation.

UR - http://www.scopus.com/inward/record.url?scp=20044391979&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20044391979&partnerID=8YFLogxK

U2 - 10.1074/jbc.M411224200

DO - 10.1074/jbc.M411224200

M3 - Article

VL - 280

SP - 5781

EP - 5788

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 7

ER -