Involvement of gap junctions between smooth muscle cells in sustained hypoxic pulmonary vasoconstriction development: A potential role for 15-HETE and 20-HETE

Igor V. Kizub, Anand Lakhkar, Vidhi Dhagia, Sachindra R. Joshi, Houli Jiang, Michael S. Wolin, J R Falck, Sreenivasulu Reddy Koduru, Ramu Errabelli, Elizabeth R. Jacobs, Michal L. Schwartzman, Sachin A. Gupte

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

In response to hypoxia, the pulmonary artery normally constricts to maintain optimal ventilation-perfusion matching in the lung, but chronic hypoxia leads to the development of pulmonary hypertension. The mechanisms of sustained hypoxic pulmonary vasoconstriction (HPV) remain unclear. The aim of this study was to determine the role of gap junctions (GJs) between smooth muscle cells (SMCs) in the sustained HPV development and involvement of arachidonic acid (AA) metabolites in GJ-mediated signaling. Vascular tone was measured in bovine intrapulmonary arteries (BIPAs) using isometric force measurement technique. Expression of contractile proteins was determined by Western blot. AA metabolites in the bath fluid were analyzed by mass spectrometry. Prolonged hypoxia elicited endothelium-independent sustained HPV in BIPAs. Inhibition of GJs by 18β-glycyrrhetinic acid (18β-GA) and heptanol, nonspecific blockers, and Gap-27, a specific blocker, decreased HPV in deendothelized BIPAs. The sustained HPV was not dependent on Ca2+ entry but decreased by removal of Ca2+ and by Rho-kinase inhibition with Y-27632. Furthermore, inhibition of GJs decreased smooth muscle myosin heavy chain (SM-MHC) expression and myosin light chain phosphorylation in BIPAs. Interestingly, inhibition of 15-and 20-hydroxyeicosatetraenoic acid (HETE) synthesis decreased HPV in deendothelized BIPAs. 15-HETE-and 20-HETE-stimulated constriction of BIPAs was inhibited by 18β-GA and Gap-27. Application of 15-HETE and 20-HETE to BIPAs increased SM-MHC expression, which was also suppressed by 18β-GA and by inhibitors of lipoxygenase and cytochrome P450 monooxygenases. More interestingly, 15,20-dihydroxyeicosatetraenoic acid and 20-OH-prostaglandin E2, novel derivatives of 20-HETE, were detected in tissue bath fluid and synthesis of these derivatives was almost completely abolished by 18β-GA. Taken together, our novel findings show that GJs between SMCs are involved in the sustained HPV in BIPAs, and 15-HETE and 20-HETE, through GJs, appear to mediate SM-MHC expression and contribute to the sustained HPV development.

Original languageEnglish (US)
Pages (from-to)L772-L783
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume310
Issue number8
DOIs
StatePublished - Apr 15 2016

Fingerprint

Gap Junctions
Vasoconstriction
Smooth Muscle Myocytes
Arteries
Glycyrrhetinic Acid
Lung
Smooth Muscle Myosins
Myosin Heavy Chains
Baths
Arachidonic Acid
Heptanol
20-hydroxy-5,8,11,14-eicosatetraenoic acid
Contractile Proteins
Lipoxygenase Inhibitors
rho-Associated Kinases
Myosin Light Chains
Mixed Function Oxygenases
Dinoprostone
Pulmonary Hypertension
Constriction

Keywords

  • 15-hydroxyeicosatetraenoic
  • Arachidonic acid metabolites
  • Bovine pulmonary artery
  • Gap junctions
  • Hypoxic pulmonary vasoconstriction

ASJC Scopus subject areas

  • Physiology
  • Medicine(all)
  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology (medical)

Cite this

Involvement of gap junctions between smooth muscle cells in sustained hypoxic pulmonary vasoconstriction development : A potential role for 15-HETE and 20-HETE. / Kizub, Igor V.; Lakhkar, Anand; Dhagia, Vidhi; Joshi, Sachindra R.; Jiang, Houli; Wolin, Michael S.; Falck, J R; Koduru, Sreenivasulu Reddy; Errabelli, Ramu; Jacobs, Elizabeth R.; Schwartzman, Michal L.; Gupte, Sachin A.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 310, No. 8, 15.04.2016, p. L772-L783.

Research output: Contribution to journalArticle

Kizub, Igor V. ; Lakhkar, Anand ; Dhagia, Vidhi ; Joshi, Sachindra R. ; Jiang, Houli ; Wolin, Michael S. ; Falck, J R ; Koduru, Sreenivasulu Reddy ; Errabelli, Ramu ; Jacobs, Elizabeth R. ; Schwartzman, Michal L. ; Gupte, Sachin A. / Involvement of gap junctions between smooth muscle cells in sustained hypoxic pulmonary vasoconstriction development : A potential role for 15-HETE and 20-HETE. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2016 ; Vol. 310, No. 8. pp. L772-L783.
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AU - Joshi, Sachindra R.

AU - Jiang, Houli

AU - Wolin, Michael S.

AU - Falck, J R

AU - Koduru, Sreenivasulu Reddy

AU - Errabelli, Ramu

AU - Jacobs, Elizabeth R.

AU - Schwartzman, Michal L.

AU - Gupte, Sachin A.

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N2 - In response to hypoxia, the pulmonary artery normally constricts to maintain optimal ventilation-perfusion matching in the lung, but chronic hypoxia leads to the development of pulmonary hypertension. The mechanisms of sustained hypoxic pulmonary vasoconstriction (HPV) remain unclear. The aim of this study was to determine the role of gap junctions (GJs) between smooth muscle cells (SMCs) in the sustained HPV development and involvement of arachidonic acid (AA) metabolites in GJ-mediated signaling. Vascular tone was measured in bovine intrapulmonary arteries (BIPAs) using isometric force measurement technique. Expression of contractile proteins was determined by Western blot. AA metabolites in the bath fluid were analyzed by mass spectrometry. Prolonged hypoxia elicited endothelium-independent sustained HPV in BIPAs. Inhibition of GJs by 18β-glycyrrhetinic acid (18β-GA) and heptanol, nonspecific blockers, and Gap-27, a specific blocker, decreased HPV in deendothelized BIPAs. The sustained HPV was not dependent on Ca2+ entry but decreased by removal of Ca2+ and by Rho-kinase inhibition with Y-27632. Furthermore, inhibition of GJs decreased smooth muscle myosin heavy chain (SM-MHC) expression and myosin light chain phosphorylation in BIPAs. Interestingly, inhibition of 15-and 20-hydroxyeicosatetraenoic acid (HETE) synthesis decreased HPV in deendothelized BIPAs. 15-HETE-and 20-HETE-stimulated constriction of BIPAs was inhibited by 18β-GA and Gap-27. Application of 15-HETE and 20-HETE to BIPAs increased SM-MHC expression, which was also suppressed by 18β-GA and by inhibitors of lipoxygenase and cytochrome P450 monooxygenases. More interestingly, 15,20-dihydroxyeicosatetraenoic acid and 20-OH-prostaglandin E2, novel derivatives of 20-HETE, were detected in tissue bath fluid and synthesis of these derivatives was almost completely abolished by 18β-GA. Taken together, our novel findings show that GJs between SMCs are involved in the sustained HPV in BIPAs, and 15-HETE and 20-HETE, through GJs, appear to mediate SM-MHC expression and contribute to the sustained HPV development.

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