Oxidized-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine induces vascular endothelial superoxide production: Implication of NADPH oxidase

Mahsa Rouhanizadeh, Juliana Hwang, Roza E. Clempus, Laura Marcu, Bernard Lassègue, Alex Sevanian, Tzung K. Hsiai

Research output: Contribution to journalArticle

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Abstract

Modified low-density lipoprotein (LDL) induces reactive oxygen species (ROS) production by vascular cells. It is unknown if specific oxidized components in these LDL particles such as oxidized-1-palmitoyl-2-arachidonoyl- sn-glycero-3-phosphorylcholine (ox-PAPC) can stimulate ROS production. Bovine aortic endothelial cells (BAEC) were incubated with ox-PAPC (50 μg/ml). At 4 h, ox-PAPC significantly enhanced the rate of O2 -. production. Pretreatment of BAEC in glucose-free Dulbecco's modified Eagle's medium plus 10 mM 2-deoxyglucose (2-DOG), the latter being an antimetabolite that blocks NADPH production by the pentose shunt, significantly reduced the rate of O2 -. production. The intensity of NAD(P)H autofluorescence decreased by 28 ± 12% in BAEC incubated with ox-PAPC compared to untreated cells, with a further decrease in the presence of 2-DOG. Ox-PAPC also increased Nox4 mRNA expression by 2.4-fold ± 0.1 while pretreatment of BAEC with the small interfering RNA (siNox4) attenuated Nox4 RNA expression. Ox-PAPC further reduced the level of glutathione while pretreatment with apocynin (100 μM) restored the GSH level (control = 22.54 ± 0.23, GSH = 18.06 ± 0.98, apocynin = 22.55 ± 0.60, ox-PAPC + apocynin = 21.17 ± 0.36 nmol/106 cells). Treatment with ox-PAPC also increased MMP-2 mRNA expression accompanied by a 1.5-fold increase in MMP-2 activity. Ox-PAPC induced vascular endothelial O2- . production that appears to be mediated largely by NADPH oxidase activity.

Original languageEnglish (US)
Pages (from-to)1512-1522
Number of pages11
JournalFree Radical Biology and Medicine
Volume39
Issue number11
DOIs
StatePublished - Dec 1 2005
Externally publishedYes

Fingerprint

Phosphorylcholine
NADPH Oxidase
Superoxides
Blood Vessels
Endothelial cells
Endothelial Cells
Deoxyglucose
Matrix Metalloproteinases
LDL Lipoproteins
Reactive Oxygen Species
Pentoses
Antimetabolites
Pentose Phosphate Pathway
Messenger RNA
Eagles
Level control
NADP
NAD
Small Interfering RNA
Glutathione

Keywords

  • Matrix metalloproteinases (MMPs)
  • NAD(P)H oxidase
  • Ox-PAPC
  • Superoxide

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

Oxidized-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine induces vascular endothelial superoxide production : Implication of NADPH oxidase. / Rouhanizadeh, Mahsa; Hwang, Juliana; Clempus, Roza E.; Marcu, Laura; Lassègue, Bernard; Sevanian, Alex; Hsiai, Tzung K.

In: Free Radical Biology and Medicine, Vol. 39, No. 11, 01.12.2005, p. 1512-1522.

Research output: Contribution to journalArticle

Rouhanizadeh, Mahsa ; Hwang, Juliana ; Clempus, Roza E. ; Marcu, Laura ; Lassègue, Bernard ; Sevanian, Alex ; Hsiai, Tzung K. / Oxidized-1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphorylcholine induces vascular endothelial superoxide production : Implication of NADPH oxidase. In: Free Radical Biology and Medicine. 2005 ; Vol. 39, No. 11. pp. 1512-1522.
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abstract = "Modified low-density lipoprotein (LDL) induces reactive oxygen species (ROS) production by vascular cells. It is unknown if specific oxidized components in these LDL particles such as oxidized-1-palmitoyl-2-arachidonoyl- sn-glycero-3-phosphorylcholine (ox-PAPC) can stimulate ROS production. Bovine aortic endothelial cells (BAEC) were incubated with ox-PAPC (50 μg/ml). At 4 h, ox-PAPC significantly enhanced the rate of O2 -. production. Pretreatment of BAEC in glucose-free Dulbecco's modified Eagle's medium plus 10 mM 2-deoxyglucose (2-DOG), the latter being an antimetabolite that blocks NADPH production by the pentose shunt, significantly reduced the rate of O2 -. production. The intensity of NAD(P)H autofluorescence decreased by 28 ± 12{\%} in BAEC incubated with ox-PAPC compared to untreated cells, with a further decrease in the presence of 2-DOG. Ox-PAPC also increased Nox4 mRNA expression by 2.4-fold ± 0.1 while pretreatment of BAEC with the small interfering RNA (siNox4) attenuated Nox4 RNA expression. Ox-PAPC further reduced the level of glutathione while pretreatment with apocynin (100 μM) restored the GSH level (control = 22.54 ± 0.23, GSH = 18.06 ± 0.98, apocynin = 22.55 ± 0.60, ox-PAPC + apocynin = 21.17 ± 0.36 nmol/106 cells). Treatment with ox-PAPC also increased MMP-2 mRNA expression accompanied by a 1.5-fold increase in MMP-2 activity. Ox-PAPC induced vascular endothelial O2- . production that appears to be mediated largely by NADPH oxidase activity.",
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T2 - Implication of NADPH oxidase

AU - Rouhanizadeh, Mahsa

AU - Hwang, Juliana

AU - Clempus, Roza E.

AU - Marcu, Laura

AU - Lassègue, Bernard

AU - Sevanian, Alex

AU - Hsiai, Tzung K.

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