Hyperoxia alters phorbol ester-induced phospholipase D activation in bovine lung microvascular endothelial cells

Shukla Roy, Narasimham Parinandi, Roy Zeigelstein, Qinghua Hu, Yong Pei, Jeffrey B. Travers, Viswanathan Natarajan

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

We investigated the effect of hyperoxia on phospholipase D (PLD) activation in bovine lung microvascular endothelial cells (BLMVECs). Generation of intracellular reactive oxygen species in BLMVECs exposed to byperoxia for 2 or 24 h was three-fold higher compared with normoxic cells as measured by dichlorodihydrofluorescein di(acetoxymethyl ester) fluorescence. Exposure of BLMVECs to hyperoxia for 2 or 24 h attenuated 12-O-tetradecanoylphorbol 13-acetate (TPA)-mediated PLD activation compared with normoxic cells, however, hyperoxia did not alter basal PLD activity. Antioxidants, such as propyl gallate and pyrrolidine dithiocarbamate, reversed the effect of hyperoxia on TPA-induced PLD activity. Furthermore, the TPA-induced PLD activation was inhibited not only by the protein kinase C inhibitor, Go6976, but also by the tyrosine kinase inhibitor, genistein, and by the Src kinase specific inhibitor, PP-2, suggesting the involvement of protein kinase C and also tyrosine kinases in TPA-induced PLD activition. Western blot analysis of cell lysates from the hyperoxic (2 or 24 h) BLMVECs stimulated with TPA with anti-phosphotyrosine antibody showed an attenuation in overall tyrosine phosphorylation of proteins. In conclusion, we have demonstrated that hyperoxia enhanced the generation of reactive oxygen species in lung microvascular endothelial cells and attenuated TPA-induced protein tyrosine phosphorylation and PLD activation. As protein tyrosine phosphorylation and PLD play important roles in inflammatory responses, this could provide a mechanism for the regulation of endothelial barrier function during hyperoxic lung injury.

Original languageEnglish (US)
Pages (from-to)217-228
Number of pages12
JournalAntioxidants and Redox Signaling
Volume5
Issue number2
DOIs
StatePublished - Apr 2003

Fingerprint

Phospholipase D
Hyperoxia
Endothelial cells
Phorbol Esters
Tetradecanoylphorbol Acetate
Endothelial Cells
Chemical activation
Lung
Acetates
Phosphorylation
Tyrosine
Protein-Tyrosine Kinases
Protein Kinase C
Reactive Oxygen Species
Propyl Gallate
Phosphotyrosine
Proteins
src-Family Kinases
Protein C Inhibitor
Genistein

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Hyperoxia alters phorbol ester-induced phospholipase D activation in bovine lung microvascular endothelial cells. / Roy, Shukla; Parinandi, Narasimham; Zeigelstein, Roy; Hu, Qinghua; Pei, Yong; Travers, Jeffrey B.; Natarajan, Viswanathan.

In: Antioxidants and Redox Signaling, Vol. 5, No. 2, 04.2003, p. 217-228.

Research output: Contribution to journalArticle

Roy, Shukla ; Parinandi, Narasimham ; Zeigelstein, Roy ; Hu, Qinghua ; Pei, Yong ; Travers, Jeffrey B. ; Natarajan, Viswanathan. / Hyperoxia alters phorbol ester-induced phospholipase D activation in bovine lung microvascular endothelial cells. In: Antioxidants and Redox Signaling. 2003 ; Vol. 5, No. 2. pp. 217-228.
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