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Cross-talk between NADPH oxidase-PKCα-p38MAPK and NF-κB-MT1MMP in activating proMMP-2 by ET-1 in pulmonary artery smooth muscle cells

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Abstract

Treatment of bovine pulmonary artery smooth muscle cells with endothelin-1 (ET-1) caused an increase in the expression and activation of proMMP-2 in the cells. The present study was undertaken to determine the underlying mechanisms involved in this scenario. We demonstrated that (i) pretreatment with NADPH oxidase inhibitor, apocynin; PKC-α inhibitor, Go6976; p38MAPK inhibitor SB203580 and NF-κB inhibitor, Bay11-7082 inhibited the expression and activation of proMMP-2 induced by ET-1; (ii) ET-1 treatment to the cells stimulated NADPH oxidase and PKCα activity, p38MAPK phosphorylation as well as NF-κB activation by translocation of NF-κBp65 subunit from cytosol to the nucleus, and subsequently by increasing its DNA-binding activity; (iii) ET-1 increases MT1-MMP expression, which was inhibited upon pretreatment with apocynin, Go6976, SB293580, and Bay 11-7082; (iv) ET-1 treatment to the cells downregulated TIMP-2 level. Although apocynin and Go6976 pretreatment reversed ET-1 effect on TIMP-2 level, yet pretreatment of the cells with SB203580 and Bay 11-7082 did not show any discernible change in TIMP-2 level by ET-1. Overall, our results suggest that ET-1-induced activation of proMMP-2 is mediated via cross-talk between NADPH oxidase-PKCα-p38MAPK and NFκB-MT1MMP signaling pathways along with a marked decrease in TIMP-2 expression in the cells.

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Abbreviations

SMC:

Smooth muscle cell

proMMP-2:

Pro matrix metalloprotease 2

MT1-MMP:

Membrane type 1 matrix metalloprotease

TIMP-2:

Tissue inhibitor of matrix metalloprotease 2

PKC-α:

Protein kinase C α

IKK:

Inhibitory κB kinase

NF-κB:

Nuclear factor κB, IκB-α, inhibitory κBα

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Acknowledgments

Financial assistance from the Council for Scientific and Industrial Research (CSIR, New Delhi) and University grants commission (UGC, New Delhi) are greatly acknowledged.

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Authors and Affiliations

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

    Jaganmay Sarkar, Animesh Chowdhury, Tapati Chakraborti & Sajal Chakraborti

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

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Sarkar, J., Chowdhury, A., Chakraborti, T. et al. Cross-talk between NADPH oxidase-PKCα-p38MAPK and NF-κB-MT1MMP in activating proMMP-2 by ET-1 in pulmonary artery smooth muscle cells. Mol Cell Biochem 415, 13–28 (2016). https://doi.org/10.1007/s11010-016-2673-6

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