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Urocortin-2 suppression of p38-MAPK signaling as an additional mechanism for ischemic cardioprotection

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Abstract

Urocortin-2 (UCN2) is cardioprotective in ischemia/reperfusion injury (I/R) through short-lived activation of ERK1/2. Key factors involved in I/R, e.g. apoptosis, mitochondrial damage, p38 kinase, and Bcl-2 family, have not been well-investigated in UCN2-induced cardioprotection. We assessed the role of p38-MAPK in anti-apoptotic Bcl-2 signaling and mitochondrial stabilization as a putative mechanisms in UCN2-induced cardioprotection. Isolated hearts from adult Sprague–Dawley rats and cultured H9c2 cells were subjected to I/R protocols with or without 10 nM UCN2 treatment. The effect of a specific p38 inhibitor SB202190 was tested in H9c2 cells. Cardiac function, LDH release, and mitochondrial membrane potential (MMP) were used to assess the degree of myocardial injury in hearts and H9c2 cells. Post-perfusion, hearts were collected for Western blot analyses or mitochondria/cytosol isolation to analyze p38 activation and Bcl-2 family members. UCN2 treatment improved rate-pressure product (58 ± 5 vs. 31 ± 4 % of Baseline; P < 0.05) and decreased LDH release (20 ± 9 vs. 90 ± 40 mU/ml LDH, P < 0.01) at the end of 60 min reperfusion. UCN2 reduced phospho-p38 levels and Bax activation. UCN2 increased the expression of Bcl-2 and inhibited the accumulation of p-Bim. With additional experiments, it was confirmed that UCN2 increases the phosphorylation of ERK1/2 in the early phase of UCN2 treatment and increases the overshot recovery of ERK1/2 phosphorylation during reperfusion. UCN2 and SB202190 partially prevented the loss of MMP induced by I/R. However, combined treatment with UCN2 and SB202190 did not provide additive benefit. UCN2 is cardioprotective in I/R in association with reduced phosphorylation of p38 together with the increased ERK1/2 activation and increased Bcl-2 family member pro-survival signaling. These changes may stabilize cardiac mitochondria, similar to p38 inhibitors, as part of a pro-survival mechanism during I/R.

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Acknowledgments

We appreciate the contribution of Dr. Paul Wolkowicz in the preparation of this manuscript. This work was supported by Cardiovascular Research Institute Start-up Fund (National University of Singapore, Singapore) and National Medical Research Council Center Grant (Ministry of Health, Singapore).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. Huashan Hospital, Fudan University, Shanghai, China

    Xiu-Fang Gao

  2. Cardiovascular Research Institute, National University of Singapore, Singapore, 117599, Singapore

    Yue Zhou, Kar-Sheng Lew, Arthur Mark Richards & Peipei Wang

  3. Department of Medicine, Centre for Translational Medicine, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore, 117599, Singapore

    Yue Zhou, Kar-Sheng Lew, Arthur Mark Richards & Peipei Wang

  4. Putuo Hospital, Shanghai, China

    Da-Ying Wang

  5. Department of Medicine, Christchurch Heart Institute, University of Otago-Christchurch, Christchurch, New Zealand

    Arthur Mark Richards

  6. Cardiac Department, National University Health System, Singapore, Singapore

    Arthur Mark Richards

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  1. Xiu-Fang Gao
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Correspondence to Peipei Wang.

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Xiu-Fang Gao and Yue Zhou have contributed equally to this work.

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Gao, XF., Zhou, Y., Wang, DY. et al. Urocortin-2 suppression of p38-MAPK signaling as an additional mechanism for ischemic cardioprotection. Mol Cell Biochem 398, 135–146 (2015). https://doi.org/10.1007/s11010-014-2213-1

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