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Endothelin-1 mediated glycosaminoglycan synthesizing gene expression involves NOX-dependent transactivation of the transforming growth factor-β receptor

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Abstract

G protein-coupled receptor (GPCR) agonist endothelin-1 (ET-1) through transactivation of the transforming growth factor (TGF) β receptor (TGFBR1) stimulates glycosaminoglycan (GAG) elongation on proteoglycans. GPCR agonists thrombin and lysophosphatidic acid (LPA) via respective receptors transactivate the TGFBR1 via Rho/ROCK dependent pathways however mechanistic insight for ET-1 transactivation of the TGFBR1 remains unknown. NADPH oxidase (NOX) generates reactive oxygen species (ROS) and is a signalling entity implicated in the pathogenesis of many diseases including atherosclerosis. If implicated in this pathway, NOX/ROS would be a potential therapeutic target. In this study, we investigated the involvement of NOX in ET-1/ET receptor-mediated transactivation of TGFBR1 to stimulate mRNA expression of GAG chain synthesizing enzymes chondroitin 4–O–sulfotransferase 1 (C4ST-1) and chondroitin sulfate synthase 1 (ChSy-1). The invitro model used vascular smooth muscle cells that were treated with pharmacological antagonists in the presence and absence of ET-1 or TGF-β. Proteins and phosphoproteins isolated from treated cells were quantified by western blotting and quantitative real-time PCR was used to assess mRNA expression of GAG synthesizing enzymes. In the presence of diphenyliodonium (DPI) (NOX inhibitor), ET-1 stimulated phospho-Smad2C levels were inhibited. ET-1 mediated mRNA expression of GAG synthesizing enzymes C4ST-1 and ChSy-1 was also blocked by TGBFR1 antagonists, SB431542, broad spectrum ET receptor antagonist bosentan, DPI and ROS scavenger N-acetyl-l-cysteine. This work shows that NOX and ROS play an important role in ET-1 mediated transactivation of the TGFBR1 and downstream gene targets associated with GAG chain elongation. As ROS is involved in GPCR to protein tyrosine kinase receptor transactivation, the NOX/ROS axis presents as the first common biochemical target in all GPCR to kinase receptor transactivation signalling.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by the Ahvaz Jundishapur University of Medical Sciences (Grant No. HLRC-9401). The support was received from The University of Queensland through a personal support package to PJL and by The University of Queensland Early Career Grant (DK) (Grant No. 1832825). DK is supported by NHMRC-Peter Doherty (1160925) and National Heart Foundation (102129) fellowships. RM is supported by The University of Queensland Research Training Programme (RTP) scholarship.

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medicine, Hyperlipidemia Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Hossein Babaahmadi-Rezaei, Ghorban Mohammad Zadeh, Reyhaneh Niayesh Mehr & Parisa Dayati

  2. School of Pharmacy, Pharmacy Australia Centre of Excellence, The University of Queensland, 20 Cornwall St, Woolloongabba, QLD, 4102, Australia

    Peter J. Little, Raafat Mohamed & Danielle Kamato

  3. Department of Pharmacy, Xinhua College of Sun Yat-Sen University, Tianhe District, Guangzhou, 510520, Guangdong, China

    Peter J. Little

  4. Department of Clinical Biochemistry, Faculty of Medicine, Cellular and Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran

    Alireza Kheirollah

  5. Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

    Reyhaneh Niayesh Mehr & Parisa Dayati

Authors
  1. Hossein Babaahmadi-Rezaei
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  2. Peter J. Little
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  3. Raafat Mohamed
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  4. Ghorban Mohammad Zadeh
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  5. Alireza Kheirollah
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  6. Reyhaneh Niayesh Mehr
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  7. Danielle Kamato
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  8. Parisa Dayati
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Contributions

Conceptualization, HB-R, DK, PJL; data curation, RM, GMZ, AK; data analysis and interpretation, PD, HB-R; drafting, RNM, PD; reviewing, DK, PJL. All authors have read and agreed to the published version of the manuscript.

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Correspondence to Danielle Kamato or Parisa Dayati.

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Babaahmadi-Rezaei, H., Little, P.J., Mohamed, R. et al. Endothelin-1 mediated glycosaminoglycan synthesizing gene expression involves NOX-dependent transactivation of the transforming growth factor-β receptor. Mol Cell Biochem 477, 981–988 (2022). https://doi.org/10.1007/s11010-021-04342-8

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