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Cardiac-specific overexpression of E3 ligase Nrdp1 increases ischemia and reperfusion-induced cardiac injury

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Abstract

Cardiomyocyte death is a major event of myocardial infarction. Previously, we and others have shown that E3 ligase-mediated protein turnover plays a critical role in cardiac injury. In this study, we sought to determine the role of a newly identified E3 ligase, neuregulin receptor degradation protein-1 (Nrdp1), on cardiac ischemia/reperfusion (I/R) injury. I/R injury markedly upregulated Nrdp1 expression in heart tissue. To elucidate the role of Nrdp1 in I/R-induced cardiac injury, neonatal cardiomyocytes were infected with adenoviral constructs expressing wild-type, dominant-negative Nrdp1 genes. Increased Nrdp1 expression enhanced I/R-induced cardiomyocyte apoptosis and inflammation as compared with the green fluorescent protein (GFP) control; these effects were attenuated by overexpression of a dominant-negative Nrdp1 (C34S/H36Q). Furthermore, cardiac-specific Nrdp1 overexpression in vivo in mouse significantly increased infarct size, the number of TUNEL-positive nuclei and inflammatory cells, as well as mortality, as compared with wild-type mice after I/R injury. The mechanisms underlying these effects were associated with the downregulation of an Nrdp1 substrate, ErbB3, accompanied by suppression of its downstream targets AKT, ERK1/2, and activation of p38 and JNK1/2. Together, these results provide evidence for an important role for Nrdp1 in regulating I/R-induced cardiac injury. Nrdp1 may constitute a new therapeutic target for ameliorating the I/R-induced cardiac injury.

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Acknowledgments

This work was supported by grants from China National Natural Science Funds for Distinguished Young Scholars (81025001; HL, 30888004; JD) and the Beijing high-level talents program (PHR20110507; HL).

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None.

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Authors and Affiliations

  1. Department of Pathology, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100005, China

    Yuan Zhang, Min Wang & Houzao Chen

  2. Department of Cardiology, Peking Union Medical Hospital, Beijing, 100730, China

    Yong Zeng & Quan Fang

  3. The Key Laboratory of Remodeling-Related Cardiovascular Diseases, Department of Pathology, School of Basic Medical Sciences, Capital Medical University, No. 10 Xitoutiao, You An Men, Beijing, 100069, China

    Cui Tian & Huihua Li

  4. Department of Genetics, National Research Institute for Family Planning, Beijing, 100081, China

    Xu Ma

  5. The Key Laboratory of Remodeling-Related Cardiovascular Diseases, Capital Medical University, Ministry of Education, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing Anzhen Hospital Affiliated the Capital Medical University, Beijing, 100029, China

    Lixin Jia & Jie Du

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  1. Yuan Zhang
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Corresponding authors

Correspondence to Jie Du or Huihua Li.

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Y. Zhang and Y. Zeng contributed equally to this work.

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Supplementary Table 1 (EPS 1003 kb)

395_2011_157_MOESM2_ESM.eps

ErbB3 receptor was expressed in the mouse heart. a The expression levels of ErbB3 mRNA in the hearts of wild-type (WT) and TG6 mice were detected by RT-PCR. Lane 1: sterilized distilled water used instead of DNA template; Lanes 2-5: ErbB3 used as DNA template. b Western blot analysis of endogenous ErbB3 protein levels from the hearts of WT and TG6 mice. c Left-ventricular sections from WT and TG6 mice stained with anti-IgG (left) or anti-ErbB3 (middle and right) antibody. Brown staining indicates ErbB3 expression. Bar = 50 μm (EPS 8554 kb)

395_2011_157_MOESM3_ESM.eps

Effect of Nrdp1 overexpression on other substrates of Nrdp1 after ischemia/reperfusion (I/R). Western blot analysis of protein levels of BRUCE, MyD88 and TBK1 from at the border area of the myocardium from WT and TG6 mice (n=5). A representative blot is shown for each condition (EPS 1004 kb)

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Zhang, Y., Zeng, Y., Wang, M. et al. Cardiac-specific overexpression of E3 ligase Nrdp1 increases ischemia and reperfusion-induced cardiac injury. Basic Res Cardiol 106, 371–383 (2011). https://doi.org/10.1007/s00395-011-0157-0

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  • DOI: https://doi.org/10.1007/s00395-011-0157-0

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