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Role of protein kinase C in oxidant — mediated activation of phospholipase A2 in rabbit pulmonary arterial smooth muscle cells

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Abstract

The present study was undertaken to test the hypothesis that activation of cell membrane associated protein kinase C (PKC) plays a role in stimulating cell membrane associated phospholipase A2 (PLA2) activity, and subsequent liberation of arachidonic acid (AA) under exposure of rabbit pulmonary arterial smooth muscle cells to the oxidant hydrogen peroxide (H2O2). Exposure of the smooth muscle cells to H2O2 dose-dependently stimulates [14C] AA release, and enhances the cell membrane associated PLA2 activity. Pretreatment of the cells with protein kinase C (PKC) inhibitors H7 and sphingosine prevent the cell membrane associated PLA2 activity, and AA release caused by H2O2. Treatment of the smooth muscle cells with H2O2 stimulates the cell membrane associated PKC activity. Pretreatment of the cells with an antioxidant vitamin E prevents H2O2 caused stimulation of the cell membrane associated PKC activity. The cell membrane associated PLA2 and PKC activities correlate linearly. These results suggest that H2O2 caused stimulation of the smooth muscle cell membrane associated PLA2 activity, and subsequent liberation of AA can occur through an increase in the activity of the cell membrane associated PKC. (Mol Cell Biochem122: 9–15, 1993)

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Abbreviations

AA:

Arachidonic Acid

PLA2 :

Phospholipase A2

PKC:

Protein Kinase C

PBS:

Phosphate Buffered Saline

HBPS:

Hank's Buffered Physiological Saline

HEPES:

4-(2-Hydroxyethyl)-1-Piperazine N-2-Ethanesulfonate

FCS:

Fetal Calf Serum

ATP:

Adenosine Triphosphate

H7 :

1-(5-isoquinolinesulfonyl)-2-methyl-piperazine

DMEM:

Dulbecco's Modified Eagles Medium

TCA:

Trichloroacetic Acid

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Chakraborti, S., Michael, J.R. Role of protein kinase C in oxidant — mediated activation of phospholipase A2 in rabbit pulmonary arterial smooth muscle cells. Mol Cell Biochem 122, 9–15 (1993). https://doi.org/10.1007/BF00925732

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