Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction

Istvan Czikora, Supriya Sridhar, Boris A Gorshkov, Irina B. Alieva, Anita Kasa, Joyce N Gonzales, Olena Potapenko, Umapathy N Siddaramappa, Helena Pillich, Ferenc G. Rick, Norman L. Block, Alexander Dmitriyevich Verin, Trinad Chakraborty, Michael A. Matthay, Andrew V. Schally, Rudolf Lucas

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Rationale: Antibiotic treatment of patients infected with G- or G+ bacteria promotes release of the toxins lipopolysaccharide (LPS) and pneumolysin (PLY) in their lungs. Growth Hormone-releasing Hormone (GHRH) agonist JI-34 protects human lung microvascular endothelial cells (HL-MVEC), expressing splice variant 1 (SV-1) of the receptor, from PLY-induced barrier dysfunction. We investigated whether JI-34 also blunts LPS-induced hyperpermeability. Since GHRH receptor (GHRH-R) signaling can potentially stimulate both cAMP-dependent barrier-protective pathways as well as barrier-disruptive protein kinase C pathways, we studied their interaction in GHRH agonist-treated HL-MVEC, in the presence of PLY, by means of siRNA-mediated protein kinase A (PKA) depletion. Methods: Barrier function measurements were done in HL-MVEC monolayers using Electrical Cell substrate Impedance Sensing (ECIS) and VE-cadherin expression by Western blotting. Capillary leak was assessed by Evans Blue dye (EBD) incorporation. Cytokine generation in broncho-alveolar lavage fluid (BALF) was measured by multiplex analysis. PKA and PKC-a activity were assessed by Western blotting. Results: GHRH agonist JI-34 significantly blunts LPS-induced barrier dysfunction, at least in part by preserving VE-cadherin expression, while not affecting inflammation. In addition to activating PKA, GHRH agonist also increases PKC-α activity in PLY-treated HL-MVEC. Treatment with PLY significantly decreases resistance in control siRNA-treated HL-MVEC, but does so even more in PKA-depleted monolayers. Pretreatment with GHRH agonist blunts PLY-induced permeability in control siRNA-treated HL-MVEC, but fails to improve barrier function in PKA-depleted PLY-treated monolayers. Conclusions: GHRH signaling in HL-MVEC protects from both LPS and PLY-mediated endothelial barrier dysfunction and concurrently induces a barrier-protective PKA-mediated and a barrier-disruptive PKC-a-induced pathway in the presence of PLY, the former of which dominates the latter.

Original languageEnglish (US)
Article number00259
JournalFrontiers in Physiology
Volume5 JUL
DOIs
StatePublished - Jan 1 2014

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Bacterial Toxins
Growth Hormone-Releasing Hormone
Cyclic AMP-Dependent Protein Kinases
Lung
Endothelial Cells
Lipopolysaccharides
Small Interfering RNA
Western Blotting
Streptococcus pneumoniae plY protein
Evans Blue
Bronchoalveolar Lavage Fluid
Electric Impedance
Protein Kinase C
Permeability
Coloring Agents
Cytokines
Anti-Bacterial Agents
Inflammation
Bacteria

Keywords

  • Capillary leak
  • Growth hormone-releasing hormone
  • Lipopolysaccharide
  • Pneumolysin
  • Protein kinase A
  • Protein kinase C

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction. / Czikora, Istvan; Sridhar, Supriya; Gorshkov, Boris A; Alieva, Irina B.; Kasa, Anita; Gonzales, Joyce N; Potapenko, Olena; Siddaramappa, Umapathy N; Pillich, Helena; Rick, Ferenc G.; Block, Norman L.; Verin, Alexander Dmitriyevich; Chakraborty, Trinad; Matthay, Michael A.; Schally, Andrew V.; Lucas, Rudolf.

In: Frontiers in Physiology, Vol. 5 JUL, 00259, 01.01.2014.

Research output: Contribution to journalArticle

Czikora, I, Sridhar, S, Gorshkov, BA, Alieva, IB, Kasa, A, Gonzales, JN, Potapenko, O, Siddaramappa, UN, Pillich, H, Rick, FG, Block, NL, Verin, AD, Chakraborty, T, Matthay, MA, Schally, AV & Lucas, R 2014, 'Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction', Frontiers in Physiology, vol. 5 JUL, 00259. https://doi.org/10.3389/fphys.2014.00259
Czikora, Istvan ; Sridhar, Supriya ; Gorshkov, Boris A ; Alieva, Irina B. ; Kasa, Anita ; Gonzales, Joyce N ; Potapenko, Olena ; Siddaramappa, Umapathy N ; Pillich, Helena ; Rick, Ferenc G. ; Block, Norman L. ; Verin, Alexander Dmitriyevich ; Chakraborty, Trinad ; Matthay, Michael A. ; Schally, Andrew V. ; Lucas, Rudolf. / Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction. In: Frontiers in Physiology. 2014 ; Vol. 5 JUL.
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title = "Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction",
abstract = "Rationale: Antibiotic treatment of patients infected with G- or G+ bacteria promotes release of the toxins lipopolysaccharide (LPS) and pneumolysin (PLY) in their lungs. Growth Hormone-releasing Hormone (GHRH) agonist JI-34 protects human lung microvascular endothelial cells (HL-MVEC), expressing splice variant 1 (SV-1) of the receptor, from PLY-induced barrier dysfunction. We investigated whether JI-34 also blunts LPS-induced hyperpermeability. Since GHRH receptor (GHRH-R) signaling can potentially stimulate both cAMP-dependent barrier-protective pathways as well as barrier-disruptive protein kinase C pathways, we studied their interaction in GHRH agonist-treated HL-MVEC, in the presence of PLY, by means of siRNA-mediated protein kinase A (PKA) depletion. Methods: Barrier function measurements were done in HL-MVEC monolayers using Electrical Cell substrate Impedance Sensing (ECIS) and VE-cadherin expression by Western blotting. Capillary leak was assessed by Evans Blue dye (EBD) incorporation. Cytokine generation in broncho-alveolar lavage fluid (BALF) was measured by multiplex analysis. PKA and PKC-a activity were assessed by Western blotting. Results: GHRH agonist JI-34 significantly blunts LPS-induced barrier dysfunction, at least in part by preserving VE-cadherin expression, while not affecting inflammation. In addition to activating PKA, GHRH agonist also increases PKC-α activity in PLY-treated HL-MVEC. Treatment with PLY significantly decreases resistance in control siRNA-treated HL-MVEC, but does so even more in PKA-depleted monolayers. Pretreatment with GHRH agonist blunts PLY-induced permeability in control siRNA-treated HL-MVEC, but fails to improve barrier function in PKA-depleted PLY-treated monolayers. Conclusions: GHRH signaling in HL-MVEC protects from both LPS and PLY-mediated endothelial barrier dysfunction and concurrently induces a barrier-protective PKA-mediated and a barrier-disruptive PKC-a-induced pathway in the presence of PLY, the former of which dominates the latter.",
keywords = "Capillary leak, Growth hormone-releasing hormone, Lipopolysaccharide, Pneumolysin, Protein kinase A, Protein kinase C",
author = "Istvan Czikora and Supriya Sridhar and Gorshkov, {Boris A} and Alieva, {Irina B.} and Anita Kasa and Gonzales, {Joyce N} and Olena Potapenko and Siddaramappa, {Umapathy N} and Helena Pillich and Rick, {Ferenc G.} and Block, {Norman L.} and Verin, {Alexander Dmitriyevich} and Trinad Chakraborty and Matthay, {Michael A.} and Schally, {Andrew V.} and Rudolf Lucas",
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TY - JOUR

T1 - Protective effect of Growth Hormone-Releasing Hormone agonist in bacterial toxin-induced pulmonary barrier dysfunction

AU - Czikora, Istvan

AU - Sridhar, Supriya

AU - Gorshkov, Boris A

AU - Alieva, Irina B.

AU - Kasa, Anita

AU - Gonzales, Joyce N

AU - Potapenko, Olena

AU - Siddaramappa, Umapathy N

AU - Pillich, Helena

AU - Rick, Ferenc G.

AU - Block, Norman L.

AU - Verin, Alexander Dmitriyevich

AU - Chakraborty, Trinad

AU - Matthay, Michael A.

AU - Schally, Andrew V.

AU - Lucas, Rudolf

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Rationale: Antibiotic treatment of patients infected with G- or G+ bacteria promotes release of the toxins lipopolysaccharide (LPS) and pneumolysin (PLY) in their lungs. Growth Hormone-releasing Hormone (GHRH) agonist JI-34 protects human lung microvascular endothelial cells (HL-MVEC), expressing splice variant 1 (SV-1) of the receptor, from PLY-induced barrier dysfunction. We investigated whether JI-34 also blunts LPS-induced hyperpermeability. Since GHRH receptor (GHRH-R) signaling can potentially stimulate both cAMP-dependent barrier-protective pathways as well as barrier-disruptive protein kinase C pathways, we studied their interaction in GHRH agonist-treated HL-MVEC, in the presence of PLY, by means of siRNA-mediated protein kinase A (PKA) depletion. Methods: Barrier function measurements were done in HL-MVEC monolayers using Electrical Cell substrate Impedance Sensing (ECIS) and VE-cadherin expression by Western blotting. Capillary leak was assessed by Evans Blue dye (EBD) incorporation. Cytokine generation in broncho-alveolar lavage fluid (BALF) was measured by multiplex analysis. PKA and PKC-a activity were assessed by Western blotting. Results: GHRH agonist JI-34 significantly blunts LPS-induced barrier dysfunction, at least in part by preserving VE-cadherin expression, while not affecting inflammation. In addition to activating PKA, GHRH agonist also increases PKC-α activity in PLY-treated HL-MVEC. Treatment with PLY significantly decreases resistance in control siRNA-treated HL-MVEC, but does so even more in PKA-depleted monolayers. Pretreatment with GHRH agonist blunts PLY-induced permeability in control siRNA-treated HL-MVEC, but fails to improve barrier function in PKA-depleted PLY-treated monolayers. Conclusions: GHRH signaling in HL-MVEC protects from both LPS and PLY-mediated endothelial barrier dysfunction and concurrently induces a barrier-protective PKA-mediated and a barrier-disruptive PKC-a-induced pathway in the presence of PLY, the former of which dominates the latter.

AB - Rationale: Antibiotic treatment of patients infected with G- or G+ bacteria promotes release of the toxins lipopolysaccharide (LPS) and pneumolysin (PLY) in their lungs. Growth Hormone-releasing Hormone (GHRH) agonist JI-34 protects human lung microvascular endothelial cells (HL-MVEC), expressing splice variant 1 (SV-1) of the receptor, from PLY-induced barrier dysfunction. We investigated whether JI-34 also blunts LPS-induced hyperpermeability. Since GHRH receptor (GHRH-R) signaling can potentially stimulate both cAMP-dependent barrier-protective pathways as well as barrier-disruptive protein kinase C pathways, we studied their interaction in GHRH agonist-treated HL-MVEC, in the presence of PLY, by means of siRNA-mediated protein kinase A (PKA) depletion. Methods: Barrier function measurements were done in HL-MVEC monolayers using Electrical Cell substrate Impedance Sensing (ECIS) and VE-cadherin expression by Western blotting. Capillary leak was assessed by Evans Blue dye (EBD) incorporation. Cytokine generation in broncho-alveolar lavage fluid (BALF) was measured by multiplex analysis. PKA and PKC-a activity were assessed by Western blotting. Results: GHRH agonist JI-34 significantly blunts LPS-induced barrier dysfunction, at least in part by preserving VE-cadherin expression, while not affecting inflammation. In addition to activating PKA, GHRH agonist also increases PKC-α activity in PLY-treated HL-MVEC. Treatment with PLY significantly decreases resistance in control siRNA-treated HL-MVEC, but does so even more in PKA-depleted monolayers. Pretreatment with GHRH agonist blunts PLY-induced permeability in control siRNA-treated HL-MVEC, but fails to improve barrier function in PKA-depleted PLY-treated monolayers. Conclusions: GHRH signaling in HL-MVEC protects from both LPS and PLY-mediated endothelial barrier dysfunction and concurrently induces a barrier-protective PKA-mediated and a barrier-disruptive PKC-a-induced pathway in the presence of PLY, the former of which dominates the latter.

KW - Capillary leak

KW - Growth hormone-releasing hormone

KW - Lipopolysaccharide

KW - Pneumolysin

KW - Protein kinase A

KW - Protein kinase C

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