Differential effect of MLC kinase in TNF-α-induced endothelial cell apoptosis and barrier dysfunction

Irina Petrache, Alexander D. Verin, Michael T. Crow, Anna Birukova, Feng Liu, Joe GN Garcia

Research output: Contribution to journalArticle

186 Citations (Scopus)

Abstract

Tumor necrosis factor (TNF)-α is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-α induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-α also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-α significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-α-induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-α-induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-α-mediated apoptosis, whereas TNF-α-induced vascular permeability appears to evolve independently of contractile tension generation.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume280
Issue number6 24-6
StatePublished - 2001
Externally publishedYes

Fingerprint

Myosin-Light-Chain Kinase
Endothelial Cells
Tumor Necrosis Factor-alpha
Apoptosis
Stress Fibers
Myosin Light Chains
Cytoskeleton
Electric Impedance
Blood Vessels
Phosphorylation
Actomyosin
rho-Associated Kinases
Lung
Caspase Inhibitors
Caspase 8
Cholera Toxin
Capillary Permeability
Ketones
Permeability

Keywords

  • Acute lung injury
  • Caspases
  • Cytoskeleton
  • Permeability
  • Rho kinase
  • Tumor necrosis factor-α

ASJC Scopus subject areas

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

Cite this

Differential effect of MLC kinase in TNF-α-induced endothelial cell apoptosis and barrier dysfunction. / Petrache, Irina; Verin, Alexander D.; Crow, Michael T.; Birukova, Anna; Liu, Feng; Garcia, Joe GN.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 280, No. 6 24-6, 2001.

Research output: Contribution to journalArticle

@article{4569b60c2ce64816bd32bbea403f09be,
title = "Differential effect of MLC kinase in TNF-α-induced endothelial cell apoptosis and barrier dysfunction",
abstract = "Tumor necrosis factor (TNF)-α is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-α induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-α also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-α significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-α-induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-α-induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-α-mediated apoptosis, whereas TNF-α-induced vascular permeability appears to evolve independently of contractile tension generation.",
keywords = "Acute lung injury, Caspases, Cytoskeleton, Permeability, Rho kinase, Tumor necrosis factor-α",
author = "Irina Petrache and Verin, {Alexander D.} and Crow, {Michael T.} and Anna Birukova and Feng Liu and Garcia, {Joe GN}",
year = "2001",
language = "English (US)",
volume = "280",
journal = "American Journal of Physiology",
issn = "0363-6143",
publisher = "American Physiological Society",
number = "6 24-6",

}

TY - JOUR

T1 - Differential effect of MLC kinase in TNF-α-induced endothelial cell apoptosis and barrier dysfunction

AU - Petrache, Irina

AU - Verin, Alexander D.

AU - Crow, Michael T.

AU - Birukova, Anna

AU - Liu, Feng

AU - Garcia, Joe GN

PY - 2001

Y1 - 2001

N2 - Tumor necrosis factor (TNF)-α is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-α induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-α also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-α significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-α-induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-α-induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-α-mediated apoptosis, whereas TNF-α-induced vascular permeability appears to evolve independently of contractile tension generation.

AB - Tumor necrosis factor (TNF)-α is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-α induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-α also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethyl-ketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-α significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-α-induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-α-induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-α-mediated apoptosis, whereas TNF-α-induced vascular permeability appears to evolve independently of contractile tension generation.

KW - Acute lung injury

KW - Caspases

KW - Cytoskeleton

KW - Permeability

KW - Rho kinase

KW - Tumor necrosis factor-α

UR - http://www.scopus.com/inward/record.url?scp=0034988809&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034988809&partnerID=8YFLogxK

M3 - Article

C2 - 11350795

AN - SCOPUS:0034988809

VL - 280

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0363-6143

IS - 6 24-6

ER -