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Zinc Dyshomeostasis in Cardiomyocytes after Acute Hypoxia/Reoxygenation

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Abstract

Zinc dyshomeostasis may play a role in the pathogenesis of myocardial ischemia/reperfusion injury. The objective of this study was to investigate the expression profile of zinc regulated transporter like- and iron-regulated transporter-like proteins (ZIPs) and zinc transporter proteins (ZnTs) in cardiomyocytes and their modulation in response to hypoxia and reoxygenation. Adult rat ventricular myocytes (ARVMs) were subjected to 6 h of hypoxia, followed by 18 h of reoxygenation. Intracellular and extracellular zinc concentrations were determined using Fluozin-3 and Newport Green fluorescence, respectively. Expression of ZnTs 1, 2, 5, and 9 along with ZIPs 1, 2, 3, 6, 7, 9, 10, 11, 13, and 14 was detectable in the cardiomyocytes by real-time reverse transcriptase polymerase chain reaction. Hypoxia elicited accumulation of intracellular free zinc, but subsequent reoxygenation resulted in striking loss of intracellular free zinc and decreased the cardiomyocyte viability. Concomitantly, extracellular zinc levels dropped rapidly during hypoxia, but increased after reoxygenation. Immunoblotting analysis revealed that hypoxia increased the expression of ZnT1, but reoxygenation significantly increased the expression of ZnTs 2 and 5. Neither hypoxia nor reoxygenation altered the levels of ZnT9. Increased intracellular zinc at the end of hypoxia is related to enhanced expression of ZIPs, whereas decreased intracellular zinc during reoxygenation appears to be due to lowered expression of all ZIPs, in addition to elevated levels of ZnTs 2 and 5. These results thus suggest that there is impaired accumulation of intracellular zinc during reoxygenation, due to overexpression of specific ZnTs and downregulation of ZIP expression.

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Acknowledgements

This research was supported by grants from DST-SERB, Govt. of India (No. SB/YS/LS-222/2013), University Grants Commission (F. No.4-5(28)/2013(BSR) (FRP)) to SB.

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  1. Institute of Genetics & Hospital for Genetic Diseases, Begumpet, Osmania University, Hyderabad, Telangana, 500016, India

    Vijaya Lakshmi Bodiga & Sita Mahalaxmi Kovur

  2. Department of Biochemistry, Kakatiya University, Warangal, Telangana, 506009, India

    Sandhya Thokala & Sreedhar Bodiga

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  1. Vijaya Lakshmi Bodiga
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This investigation was approved by the Institutional Animal Ethics Committee, Kakatiya University, registered under “Committee for the Purpose of Control and Supervision of Experiments on Laboratory Animals,” Ministry of Environment and Forests, Government of India, and conforms with the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health Publication 85–23, revised 1996.

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Bodiga, V.L., Thokala, S., Kovur, S.M. et al. Zinc Dyshomeostasis in Cardiomyocytes after Acute Hypoxia/Reoxygenation. Biol Trace Elem Res 179, 117–129 (2017). https://doi.org/10.1007/s12011-017-0957-7

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