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Cross-talk between p38MAPK and Giα in regulating cPLA2 activity by ET-1 in pulmonary smooth muscle cells

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Abstract

Endothelin-1 (ET-1) is known as the most potent vasoconstrictor yet described. Infusion of ET-1 into isolated rabbit lung has been shown to cause pulmonary vasoconstriction with the involvement of arachidonic acid metabolites. Given the potency of arachidonic acid metabolites, the activity of phospholipase A2 must be tightly regulated. Herein, we determined the mechanisms by which ET-1 stimulates cPLA2 activity during ET-1 stimulation of bovine pulmonary artery smooth muscle cells. We demonstrated that (i) treatment of bovine pulmonary artery smooth muscle cells with ET-1 stimulates cPLA2 activity in the cell membrane; (ii) ET-1 caused increase in O ·−2 production occurs via NADPH oxidase-dependent mechanism; (iii) ET-1-stimulated NADPH oxidase activity is markedly prevented upon pretreatment with PKC-ζ inhibitor, indicating that PKC-ζ plays a prominent role in this scenario; (iv) ET-1-induced NADPH oxidase-derived O ·−2 stimulates an aprotinin sensitive protease activity due to prominent increase in [Ca2+]i; (v) the aprotinin sensitive protease plays a pivotal role in activating PKC-α, which in turn phosphorylates p38MAPK and subsequently Giα leading to the activation of cPLA2. Taken together, we suggest that cross-talk between p38MAPK and Giα with the involvement of PKC-ζ, NADPH oxidase-derived O ·−2 , [Ca2+]i, aprotinin-sensitive protease and PKC-α play a pivotal role for full activation of cPLA2 during ET-1 stimulation of pulmonary artery smooth muscle cells.

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Abbreviations

cPLA2 :

Cytosolic phospholipase A2

ET-1:

Endothelin-1

Giα:

α-Subunit of the inhibitory G protein

[Ca2+]i :

Intracellular Ca2+

MAPK:

Mitogen activated protein kinase

PKC:

Protein kinase C

PLC:

Phospholipase C

O ·−2 :

Superoxide anion

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Acknowledgments

Thanks are due to late Prof. Gail H Gurtner (Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland) and Prof. John R Michael (Department of Medicine, University of Utah Health Sciences Centre, Salt Lake City, Utah) for some help and suggestion. Thanks are due to Indian Council of Medical Research, New Delhi and the DST-PURSE programme of the University of Kalyani for partly financing the research. This article is dedicated to Prof. John R Michael and the late Professor Gail H Gurtner.

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  1. Department of Biochemistry and Biophysics, University of Kalyani, Kalyani, 741235, West Bengal, India

    Sajal Chakraborti, Animesh Chowdhury & Tapati Chakraborti

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  1. Sajal Chakraborti
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  2. Animesh Chowdhury
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Correspondence to Sajal Chakraborti.

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Chakraborti, S., Chowdhury, A. & Chakraborti, T. Cross-talk between p38MAPK and Giα in regulating cPLA2 activity by ET-1 in pulmonary smooth muscle cells. Mol Cell Biochem 400, 107–123 (2015). https://doi.org/10.1007/s11010-014-2267-0

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