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Downregulation of RACK1 is associated with cardiomyocyte apoptosis after myocardial ischemia/reperfusion injury in adult rats

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Abstract

The receptor for activated C kinase 1 (RACK1) is a multifaceted scaffolding protein that mediates the shuttling of activated protein kinase C (PKC) to cellular membranes. In addition, RACK1 could decrease cell apoptosis in a variety of disease models. However, the function of RACK1 in cardiomyocyte apoptosis after myocardial ischemia/reperfusion (I/R) is unknown. In this study, male Sprague–Dawley rats were anesthetized and subjected to myocardial I/R insult consisting of 30 min left anterior descending coronary artery (LAD) occlusion followed by reperfusion for 1, 2, 4, 6, 8, 12, and 24 h. The expression of RACK1 was decreased after myocardial I/R and was associated with cardiomyocyte apoptosis. To further verify the relationship between RACK1 and cardiomyocyte apoptosis, H9c2 cardiomyocytes were cultured under hypoxia for 6 h, then maintained in the regular incubator to reoxygenation. After H9c2 cells were transfected with Flag-RACK1 to overexpress RACK1, RACK1 expression was upregulated in hypoxia/reoxygenation (H/R) 4 h group accompanied with the decrease of cleaved caspase-3 and the increase of Bcl-2 expression. Terminal transferase-mediated biotin dUTP nick end labeling (TUNEL) assay showed that RACK1 overexpression inhibited H9c2 cell apoptosis induced by H/R treatment. Our data suggested that RACK1 might suppress cardiomyocyte apoptosis after I/R, providing a novel molecular target for the therapy of ischemia heart disease.

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Acknowledgments

This work was supported in part by the National Natural Science Foundation of China (Nos. 81401365, 81373223, 81200918, and 81172879), a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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    Authors and Affiliations

    1. Department of Thoracic Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, 226001, People’s Republic of China

      Long Qian, Jiahai Shi, Jiawei Lu, Xiaoning Lu, Kunpeng Wu, Chen Yang, Daliang Yan & Qingsheng You

    2. Department of Vasculocardiology, Affiliated Hospital of Nantong University, Nantong, Jiangsu, 226001, People’s Republic of China

      Chi Zhang & Chao Zhang

    3. Department of Pathogen Biology, Medical College, Nantong University, Nantong, Jiangsu, 226001, People’s Republic of China

      Xiaojuan Liu

    4. Jiangsu Province Key Laboratory for Inflammation and Molecular Drug Target, Medical College, Nantong University, Nantong, Jiangsu, 226001, People’s Republic of China

      Long Qian, Jiahai Shi, Chi Zhang, Jiawei Lu, Xiaoning Lu, Kunpeng Wu, Chen Yang, Daliang Yan, Chao Zhang, Qingsheng You & Xiaojuan Liu

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    1. Long Qian
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    Correspondence to Qingsheng You or Xiaojuan Liu.

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    Editor: Tetsuji Okamoto

    Long Qian, Jiahai Shi, Qingsheng You and Xiaojuan Liu contributed equally to this work.

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    Qian, L., Shi, J., Zhang, C. et al. Downregulation of RACK1 is associated with cardiomyocyte apoptosis after myocardial ischemia/reperfusion injury in adult rats. In Vitro Cell.Dev.Biol.-Animal 52, 305–313 (2016). https://doi.org/10.1007/s11626-015-9981-0

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