Role of transglutaminase 2 in A1 adenosine receptor- and β2 -adrenoceptor-mediated pharmacological pre- and post-conditioning against hypoxia-reoxygenation-induced cell death in H9c2 cells

Vyas, F.S., Nelson, C.P. ORCID: 0000-0003-1034-140X and Dickenson, J.M. ORCID: 0000-0002-9683-969X, 2018. Role of transglutaminase 2 in A1 adenosine receptor- and β2 -adrenoceptor-mediated pharmacological pre- and post-conditioning against hypoxia-reoxygenation-induced cell death in H9c2 cells. European Journal of Pharmacology, 819, pp. 144-160. ISSN 0014-2999

[img]
Preview
Text
Pubsub10237_Dickenson.pdf - Post-print

Download (2MB) | Preview

Abstract

Pharmacologically-induced pre- and post-conditioning represent attractive therapeutic strategies to reduce ischaemia/reperfusion injury during cardiac surgery and following myocardial infarction. We have previously reported that transglutaminase 2 (TG2) activity is modulated by the A1 adenosine receptor and β2-adrenoceptor in H9c2 cardiomyoblasts. The primary aim of this study was to determine the role of TG2 in A1 adenosine receptor and β2-adrenoceptor-induced pharmacological pre- and post-conditioning in the H9c2 cells. H9c2 cells were exposed to 8 h hypoxia (1% O2) followed by 18 h reoxygenation, after which cell viability was assessed by monitoring mitochondrial reduction of MTT, lactate dehydrogenase release and caspase-3 activation. N6-cyclopentyladenosine (CPA; A1 adenosine receptor agonist), formoterol (β2-adrenoceptor agonist) or isoprenaline (non-selective β-adrenoceptor agonist) were added before hypoxia/reoxygenation (pre-conditioning) or at the start of reoxygenation following hypoxia (post-conditioning). Pharmacological pre- and post-conditioning with CPA and isoprenaline significantly reduced hypoxia/reoxygenation-induced cell death. In contrast, formoterol did not elicit protection. Pre-treatment with pertussis toxin (Gi/o-protein inhibitor), DPCPX (A1 adenosine receptor antagonist) or TG2 inhibitors (Z-DON and R283) attenuated the A1 adenosine receptor-induced pharmacological pre- and post-conditioning. Similarly, pertussis toxin, ICI 118,551 (β2-adrenoceptor antagonist) or TG2 inhibition attenuated the isoprenaline-induced cell survival. Knockdown of TG2 using small interfering RNA (siRNA) attenuated CPA and isoprenaline-induced pharmacological pre- and post-conditioning. Finally, proteomic analysis following isoprenaline treatment identified known (e.g. protein S100-A6) and novel (e.g. adenine phosphoribosyltransferase) protein substrates for TG2. These results have shown that A1 adenosine receptor and β2-adrenoceptor-induced protection against simulated hypoxia/reoxygenation occurs in a TG2 and Gi/o-protein dependent manner in H9c2 cardiomyoblasts.

Item Type: Journal article
Publication Title: European Journal of Pharmacology
Creators: Vyas, F.S., Nelson, C.P. and Dickenson, J.M.
Publisher: Elsevier
Date: 15 January 2018
Volume: 819
ISSN: 0014-2999
Identifiers:
NumberType
10.1016/j.ejphar.2017.11.049DOI
S001429991730780XPublisher Item Identifier
Divisions: Schools > School of Science and Technology
Depositing User: Linda Sullivan
Date Added: 19 Feb 2018 09:16
Last Modified: 05 Dec 2018 03:00
URI: http://irep.ntu.ac.uk/id/eprint/32738

Actions (login required)

Edit View Edit View

Views

Views per month over past year

Downloads

Downloads per month over past year