Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91 phox to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock

Bahar Tunctan, Belma Korkmaz, Ayse Nihal Sari, Meltem Kacan, Demet Unsal, Mehmet Sami Serin, C. Kemal Buharalioglu, Seyhan Sahan-Firat, Tuba Cuez, Wolf Hagen Schunck, Vijaya L. Manthati, John R. Falck, Kafait U. Malik

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20- HETE), N-[20-hydroxyeicosa-5(Z),14(Z)- dienoyl]glycine (5,14-HEDGE), prevents vascular hyporeactivity, hypotension, tachycardia, and inflammation in rats treated with lipopolysaccharide (LPS) and mortality in endotoxemic mice. These changes were attributed to decreased production of inducible nitric oxide (NO) synthase (iNOS)-derived NO, cyclooxygenase (COX)-2-derived vasodilator prostanoids, and proinflammatory mediators associated with increased cyctochrome P450 (CYP) 4A1-derived 20-HETE and CYP2C23-dependent antiinflammatory mediator formation. The aim of this study was to determine whether decreased expression and activity of iNOS, soluble guanylyl cyclase (sGC), protein kinase G (PKG), COX-2, gp91phox (NOX2; a superoxide generating NOX enzyme), and peroxynitrite production associated with increased expression of COX-1 and CYP4A1 and 20-HETE formation in renal and cardiovascular tissues of rats contributes to the effect of 5,14-HEDGE to prevent vasodilation, hypotension, tachycardia, and inflammation in response to systemic administration of LPS. Mean arterial pressure fell by 28 mmHg and heart rate rose by 47 beats/min in LPS (10 mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of iNOS and COX-2 associated with a decrease in COX-1 and CYP4A1 mRNA and protein expression. Increased NOS activity, iNOS-heat shock protein 90 complex formation (an index for iNOS activity), protein expression of phosphorylated vasodilator stimulated phosphoprotein (an index for PKG activity), gp91phox, p47phox (NOXO2; organizer subunit of gp91phox), and nitrotyrosine (an index for peroxynitrite production) as well as cGMP (an index for sGC activity), 6-keto-PGF (a stable metabolite PGI 2) and PGE2 levels (indexes for COX activity), and nitrotyrosine levels by LPS were also associated with decreased CYP hydroxylase activity as measured by 20-HETE formation from arachidonic acid in renal microsomes of LPS-treated rats. These effects of LPS, except iNOS mRNA and COX-1 protein expression, were prevented by 5,14-HEDGE (30 mg/kg, s.c.; 1 h after LPS). A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (30 mg/kg, s.c.; 1 h after LPS) reversed the effects of 5,14-HEDGE, except iNOS and COX-1 mRNA and protein expression as well as expression of CYP4A1 mRNA. These results suggest that increased CYP4A1 expression and 20-HETE formation associated with suppression of iNOS/sGC/PKG pathway, COX-2, and gp91phox participate in the protective effect of 5,14-HEDGE against vasodilation, hypotension, tachycardia, and inflammation in the rat model of septic shock.

Original languageEnglish (US)
Pages (from-to)18-41
Number of pages24
JournalNitric Oxide - Biology and Chemistry
Volume33
DOIs
StatePublished - 2013

Fingerprint

Cyclic GMP-Dependent Protein Kinases
Guanylate Cyclase
Cyclooxygenase 2
Septic Shock
Tachycardia
Vasodilation
Hypotension
Lipopolysaccharides
Rats
Inflammation
Cyclooxygenase 1
Messenger RNA
Peroxynitrous Acid
Proteins
Kidney
HSP90 Heat-Shock Proteins
4-ethoxymethylene-2-phenyl-2-oxazoline-5-one
Soluble Guanylyl Cyclase
20-hydroxy-5,8,11,14-eicosatetraenoic acid
N-(20-hydroxyeicosa-5,14-dienoyl)glycine

Keywords

  • COX-2
  • CYP4A1
  • Endotoxin
  • Hypotension
  • INOS/sGC/PKG pathway

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Cancer Research
  • Physiology

Cite this

Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91 phox to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock. / Tunctan, Bahar; Korkmaz, Belma; Sari, Ayse Nihal; Kacan, Meltem; Unsal, Demet; Serin, Mehmet Sami; Buharalioglu, C. Kemal; Sahan-Firat, Seyhan; Cuez, Tuba; Schunck, Wolf Hagen; Manthati, Vijaya L.; Falck, John R.; Malik, Kafait U.

In: Nitric Oxide - Biology and Chemistry, Vol. 33, 2013, p. 18-41.

Research output: Contribution to journalArticle

Tunctan, Bahar ; Korkmaz, Belma ; Sari, Ayse Nihal ; Kacan, Meltem ; Unsal, Demet ; Serin, Mehmet Sami ; Buharalioglu, C. Kemal ; Sahan-Firat, Seyhan ; Cuez, Tuba ; Schunck, Wolf Hagen ; Manthati, Vijaya L. ; Falck, John R. ; Malik, Kafait U. / Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91 phox to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock. In: Nitric Oxide - Biology and Chemistry. 2013 ; Vol. 33. pp. 18-41.
@article{6add1a5ddc0f42af890cc392e51266b9,
title = "Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91 phox to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock",
abstract = "We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20- HETE), N-[20-hydroxyeicosa-5(Z),14(Z)- dienoyl]glycine (5,14-HEDGE), prevents vascular hyporeactivity, hypotension, tachycardia, and inflammation in rats treated with lipopolysaccharide (LPS) and mortality in endotoxemic mice. These changes were attributed to decreased production of inducible nitric oxide (NO) synthase (iNOS)-derived NO, cyclooxygenase (COX)-2-derived vasodilator prostanoids, and proinflammatory mediators associated with increased cyctochrome P450 (CYP) 4A1-derived 20-HETE and CYP2C23-dependent antiinflammatory mediator formation. The aim of this study was to determine whether decreased expression and activity of iNOS, soluble guanylyl cyclase (sGC), protein kinase G (PKG), COX-2, gp91phox (NOX2; a superoxide generating NOX enzyme), and peroxynitrite production associated with increased expression of COX-1 and CYP4A1 and 20-HETE formation in renal and cardiovascular tissues of rats contributes to the effect of 5,14-HEDGE to prevent vasodilation, hypotension, tachycardia, and inflammation in response to systemic administration of LPS. Mean arterial pressure fell by 28 mmHg and heart rate rose by 47 beats/min in LPS (10 mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of iNOS and COX-2 associated with a decrease in COX-1 and CYP4A1 mRNA and protein expression. Increased NOS activity, iNOS-heat shock protein 90 complex formation (an index for iNOS activity), protein expression of phosphorylated vasodilator stimulated phosphoprotein (an index for PKG activity), gp91phox, p47phox (NOXO2; organizer subunit of gp91phox), and nitrotyrosine (an index for peroxynitrite production) as well as cGMP (an index for sGC activity), 6-keto-PGF1α (a stable metabolite PGI 2) and PGE2 levels (indexes for COX activity), and nitrotyrosine levels by LPS were also associated with decreased CYP hydroxylase activity as measured by 20-HETE formation from arachidonic acid in renal microsomes of LPS-treated rats. These effects of LPS, except iNOS mRNA and COX-1 protein expression, were prevented by 5,14-HEDGE (30 mg/kg, s.c.; 1 h after LPS). A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (30 mg/kg, s.c.; 1 h after LPS) reversed the effects of 5,14-HEDGE, except iNOS and COX-1 mRNA and protein expression as well as expression of CYP4A1 mRNA. These results suggest that increased CYP4A1 expression and 20-HETE formation associated with suppression of iNOS/sGC/PKG pathway, COX-2, and gp91phox participate in the protective effect of 5,14-HEDGE against vasodilation, hypotension, tachycardia, and inflammation in the rat model of septic shock.",
keywords = "COX-2, CYP4A1, Endotoxin, Hypotension, INOS/sGC/PKG pathway",
author = "Bahar Tunctan and Belma Korkmaz and Sari, {Ayse Nihal} and Meltem Kacan and Demet Unsal and Serin, {Mehmet Sami} and Buharalioglu, {C. Kemal} and Seyhan Sahan-Firat and Tuba Cuez and Schunck, {Wolf Hagen} and Manthati, {Vijaya L.} and Falck, {John R.} and Malik, {Kafait U.}",
year = "2013",
doi = "10.1016/j.niox.2013.05.001",
language = "English (US)",
volume = "33",
pages = "18--41",
journal = "Nitric Oxide - Biology and Chemistry",
issn = "1089-8603",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Contribution of iNOS/sGC/PKG pathway, COX-2, CYP4A1, and gp91 phox to the protective effect of 5,14-HEDGE, a 20-HETE mimetic, against vasodilation, hypotension, tachycardia, and inflammation in a rat model of septic shock

AU - Tunctan, Bahar

AU - Korkmaz, Belma

AU - Sari, Ayse Nihal

AU - Kacan, Meltem

AU - Unsal, Demet

AU - Serin, Mehmet Sami

AU - Buharalioglu, C. Kemal

AU - Sahan-Firat, Seyhan

AU - Cuez, Tuba

AU - Schunck, Wolf Hagen

AU - Manthati, Vijaya L.

AU - Falck, John R.

AU - Malik, Kafait U.

PY - 2013

Y1 - 2013

N2 - We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20- HETE), N-[20-hydroxyeicosa-5(Z),14(Z)- dienoyl]glycine (5,14-HEDGE), prevents vascular hyporeactivity, hypotension, tachycardia, and inflammation in rats treated with lipopolysaccharide (LPS) and mortality in endotoxemic mice. These changes were attributed to decreased production of inducible nitric oxide (NO) synthase (iNOS)-derived NO, cyclooxygenase (COX)-2-derived vasodilator prostanoids, and proinflammatory mediators associated with increased cyctochrome P450 (CYP) 4A1-derived 20-HETE and CYP2C23-dependent antiinflammatory mediator formation. The aim of this study was to determine whether decreased expression and activity of iNOS, soluble guanylyl cyclase (sGC), protein kinase G (PKG), COX-2, gp91phox (NOX2; a superoxide generating NOX enzyme), and peroxynitrite production associated with increased expression of COX-1 and CYP4A1 and 20-HETE formation in renal and cardiovascular tissues of rats contributes to the effect of 5,14-HEDGE to prevent vasodilation, hypotension, tachycardia, and inflammation in response to systemic administration of LPS. Mean arterial pressure fell by 28 mmHg and heart rate rose by 47 beats/min in LPS (10 mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of iNOS and COX-2 associated with a decrease in COX-1 and CYP4A1 mRNA and protein expression. Increased NOS activity, iNOS-heat shock protein 90 complex formation (an index for iNOS activity), protein expression of phosphorylated vasodilator stimulated phosphoprotein (an index for PKG activity), gp91phox, p47phox (NOXO2; organizer subunit of gp91phox), and nitrotyrosine (an index for peroxynitrite production) as well as cGMP (an index for sGC activity), 6-keto-PGF1α (a stable metabolite PGI 2) and PGE2 levels (indexes for COX activity), and nitrotyrosine levels by LPS were also associated with decreased CYP hydroxylase activity as measured by 20-HETE formation from arachidonic acid in renal microsomes of LPS-treated rats. These effects of LPS, except iNOS mRNA and COX-1 protein expression, were prevented by 5,14-HEDGE (30 mg/kg, s.c.; 1 h after LPS). A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (30 mg/kg, s.c.; 1 h after LPS) reversed the effects of 5,14-HEDGE, except iNOS and COX-1 mRNA and protein expression as well as expression of CYP4A1 mRNA. These results suggest that increased CYP4A1 expression and 20-HETE formation associated with suppression of iNOS/sGC/PKG pathway, COX-2, and gp91phox participate in the protective effect of 5,14-HEDGE against vasodilation, hypotension, tachycardia, and inflammation in the rat model of septic shock.

AB - We have previously demonstrated that a stable synthetic analog of 20-hydroxyeicosatetraenoic acid (20- HETE), N-[20-hydroxyeicosa-5(Z),14(Z)- dienoyl]glycine (5,14-HEDGE), prevents vascular hyporeactivity, hypotension, tachycardia, and inflammation in rats treated with lipopolysaccharide (LPS) and mortality in endotoxemic mice. These changes were attributed to decreased production of inducible nitric oxide (NO) synthase (iNOS)-derived NO, cyclooxygenase (COX)-2-derived vasodilator prostanoids, and proinflammatory mediators associated with increased cyctochrome P450 (CYP) 4A1-derived 20-HETE and CYP2C23-dependent antiinflammatory mediator formation. The aim of this study was to determine whether decreased expression and activity of iNOS, soluble guanylyl cyclase (sGC), protein kinase G (PKG), COX-2, gp91phox (NOX2; a superoxide generating NOX enzyme), and peroxynitrite production associated with increased expression of COX-1 and CYP4A1 and 20-HETE formation in renal and cardiovascular tissues of rats contributes to the effect of 5,14-HEDGE to prevent vasodilation, hypotension, tachycardia, and inflammation in response to systemic administration of LPS. Mean arterial pressure fell by 28 mmHg and heart rate rose by 47 beats/min in LPS (10 mg/kg, i.p.)-treated rats. Administration of LPS also increased mRNA and protein expression of iNOS and COX-2 associated with a decrease in COX-1 and CYP4A1 mRNA and protein expression. Increased NOS activity, iNOS-heat shock protein 90 complex formation (an index for iNOS activity), protein expression of phosphorylated vasodilator stimulated phosphoprotein (an index for PKG activity), gp91phox, p47phox (NOXO2; organizer subunit of gp91phox), and nitrotyrosine (an index for peroxynitrite production) as well as cGMP (an index for sGC activity), 6-keto-PGF1α (a stable metabolite PGI 2) and PGE2 levels (indexes for COX activity), and nitrotyrosine levels by LPS were also associated with decreased CYP hydroxylase activity as measured by 20-HETE formation from arachidonic acid in renal microsomes of LPS-treated rats. These effects of LPS, except iNOS mRNA and COX-1 protein expression, were prevented by 5,14-HEDGE (30 mg/kg, s.c.; 1 h after LPS). A competitive antagonist of vasoconstrictor effects of 20-HETE, 20-hydroxyeicosa-6(Z), 15(Z)-dienoic acid (30 mg/kg, s.c.; 1 h after LPS) reversed the effects of 5,14-HEDGE, except iNOS and COX-1 mRNA and protein expression as well as expression of CYP4A1 mRNA. These results suggest that increased CYP4A1 expression and 20-HETE formation associated with suppression of iNOS/sGC/PKG pathway, COX-2, and gp91phox participate in the protective effect of 5,14-HEDGE against vasodilation, hypotension, tachycardia, and inflammation in the rat model of septic shock.

KW - COX-2

KW - CYP4A1

KW - Endotoxin

KW - Hypotension

KW - INOS/sGC/PKG pathway

UR - http://www.scopus.com/inward/record.url?scp=84885109307&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84885109307&partnerID=8YFLogxK

U2 - 10.1016/j.niox.2013.05.001

DO - 10.1016/j.niox.2013.05.001

M3 - Article

VL - 33

SP - 18

EP - 41

JO - Nitric Oxide - Biology and Chemistry

JF - Nitric Oxide - Biology and Chemistry

SN - 1089-8603

ER -