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Post-conditioning protects cardiomyocytes from apoptosis via PKCε-interacting with calcium-sensing receptors to inhibit endo(sarco)plasmic reticulum–mitochondria crosstalk

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Abstract

The intracellular Ca2+ concentration ([Ca2+]i) is increased during cardiac ischemia/reperfusion injury (IRI), leading to endo(sarco)plasmic reticulum (ER) stress. Persistent ER stress, such as with the accumulation of [Ca2+]i, results in apoptosis. Ischemic post-conditioning (PC) can protect cardiomyocytes from IRI by reducing the [Ca2+]i via protein kinase C (PKC). The calcium-sensing receptor (CaR), a G protein-coupled receptor, causes the production of inositol phosphate (IP3) to increase the release of intracellular Ca2+ from the ER. This process can be negatively regulated by PKC through the phosphorylation of Thr-888 of the CaR. This study tested the hypothesis that PC prevents cardiomyocyte apoptosis by reducing the [Ca2+]i through an interaction of PKC with CaR to alleviate [Ca2+]ER depletion and [Ca2+]m elevation by the ER-mitochondrial associated membrane (MAM). Cardiomyocytes were post-conditioned after 3 h of ischemia by three cycles of 5 min of reperfusion and 5 min of re-ischemia before 6 h of reperfusion. During PC, PKCε translocated to the cell membrane and interacted with CaR. While PC led to a significant decrease in [Ca2+]i, the [Ca2+]ER was not reduced and [Ca2+]m was not increased in the PC and GdCl3–PC groups. Furthermore, there was no evident ∆ψm collapse during PC compared with ischemia/reperfusion (I/R) or PKC inhibitor groups, as evaluated by laser confocal scanning microscopy. The apoptotic rates detected by TUNEL and Hoechst33342 were lower in PC and GdCl3–PC groups than those in I/R and PKC inhibitor groups. Apoptotic proteins, including m-calpain, BAP31, and caspase-12, were significantly increased in the I/R and PKC inhibitor groups. These results suggested that PKCε interacting with CaR protected post-conditioned cardiomyocytes from programmed cell death by inhibiting disruption of the mitochondria by the ER as well as preventing calcium-induced signaling of the apoptotic pathway.

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Acknowledgments

This study was supported by grants from the National Basic Research Program of China (973 program No. 2007CB512000), and the National Natural Science Foundation of China (No. 30700288, 30770878, 30871012).

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Authors and Affiliations

  1. Department of Pathophysiology, Harbin Medical University, Harbin, 150086, China

    Shiyun Dong, Fang-hao Lu, Ya-jun Zhao, Ye Tian, Chang-qing Xu & Wei-hua Zhang

  2. Department of Geriatric, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150086, China

    Zongyan Teng

  3. Department of Neurobiology, Harbin Medical University, Harbin, 150086, China

    Hulun Li

  4. Department of Immunology, Harbin Medical University, Harbin, 150086, China

    Huan Ren

  5. Department of Pathology, Harbin Medical University, Harbin, 150086, China

    He Chen

  6. Department of Pharmacology, Harbin Medical University, Harbin, 150086, China

    Zhen-wei Pan & Yan-jie Lv

  7. Bio-Pharmaceutical Key Laboratory of Heilongjiang Province, Harbin Medical University, Harbin, 150086, China

    Bao-feng Yang, Chang-qing Xu & Wei-hua Zhang

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  1. Shiyun Dong
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Correspondence to Chang-qing Xu or Wei-hua Zhang.

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Shiyun Dong and Zongyan Teng contributed equally to this study.

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Dong, S., Teng, Z., Lu, Fh. et al. Post-conditioning protects cardiomyocytes from apoptosis via PKCε-interacting with calcium-sensing receptors to inhibit endo(sarco)plasmic reticulum–mitochondria crosstalk. Mol Cell Biochem 341, 195–206 (2010). https://doi.org/10.1007/s11010-010-0450-5

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