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Hydrogen sulfide mediates the cardioprotective effects of gene therapy with PKG-Iα

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Abstract

Cyclic GMP-dependent protein kinase (PKG) is a serine-threonine kinase that mediates the cardioprotective effect of ischemic and pharmacologic preconditioning. Since hydrogen sulfide (H2S) has been implicated in mediating the cardioprotective effects of the cGMP modulators tadalafil and cinaciguat, we tested the hypothesis that myocardial gene therapy with PKG exerts cardioprotection against ischemia/reperfusion (I/R) injury through a mechanism involving H2S. Adult rat cardiomyocytes were infected with adenoviral vector encoding PKGIα or inactive mutant PKGIαK390A (K390A) for 24 h. Necrosis and apoptosis (n = 6/group) were determined after 90 min of simulated ischemia and 1 or 18 h of reoxygenation, respectively. To study the effect of PKGIα in vivo, mice received intramyocardial injections of adenoviral PKGIα or K390A. Four days later, the hearts were subjected to 30 min of ischemia followed by reperfusion for 24 h. The inhibitor of H2S-producing enzyme, cystathionine-γ-lyase (CSE), dl-propargylglycine (PAG, 50 mg/kg, ip) was given 30 min before ischemia. PKGIα overexpression induced CSE expression, whereas cystathionine-β-synthase (CBS) and 3-mercaptopyruvate sulfurtransferase expression was not changed. PKGIα overexpression increased H2S in the heart and cardiomyocytes in relation to control and PKGIαK390A. Moreover, PAG abolished protection with PKGIα in vitro by increasing necrosis (35.2 ± 1.7 %, P < 0.05) and apoptosis (23.5 ± 1.8 %, P < 0.05) as compared to PKGIα-overexpressing cells (necrosis: 17.2 ± 0.9 % and apoptosis: 13.2 ± 0.8 %). In vivo, PKGIα overexpression reduced infarct size and preserved left ventricular fractional shortening as compared with K390A (P < 0.05) and PAG abolished the cardioprotective effect of PKGIα. The protective effect of myocardial gene therapy with PKGIα against I/R injury is mediated through a mechanism involving H2S signaling.

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Acknowledgments

The authors would like to thank Jun He, M.S. and Dr. Nitai Mukhopadhyay for performing the biostatistical analyses for this study. This study was supported in part by a grant from the American Heart Association (BGIA 0765273U) to AD, grants from the National Institutes of Health (HL51045, HL59469, and HL79424) to RCK, and grants from the American Heart Association (10SDG3770011 and 14GRNT20010003) and the Virginia Commonwealth University Presidential Research Quest Fund to FNS.

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On behalf of all authors, the corresponding author states that there is no conflict of interest.

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  1. Division of Cardiology, Department of Internal Medicine, Pauley Heart Center, Virginia Commonwealth University, Box 980204, 1101 East Marshall Street, Room 7-070, Richmond, VA, 23298-0204, USA

    Anindita Das, Arun Samidurai, Nicholas N. Hoke, Rakesh C. Kukreja & Fadi N. Salloum

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  1. Anindita Das
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  2. Arun Samidurai
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Correspondence to Fadi N. Salloum.

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Das, A., Samidurai, A., Hoke, N.N. et al. Hydrogen sulfide mediates the cardioprotective effects of gene therapy with PKG-Iα. Basic Res Cardiol 110, 42 (2015). https://doi.org/10.1007/s00395-015-0500-y

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