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Clopidogrel attenuates lithium-induced alterations in renal water and sodium channels/transporters in mice

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Abstract

Lithium (Li) administration causes deranged expression and function of renal aquaporins and sodium channels/transporters resulting in nephrogenic diabetes insipidus (NDI). Extracellular nucleotides (ATP/ADP/UTP), via P2 receptors, regulate these transport functions. We tested whether clopidogrel bisulfate (CLPD), an antagonist of ADP-activated P2Y12 receptor, would affect Li-induced alterations in renal aquaporins and sodium channels/transporters. Adult mice were treated for 14 days with CLPD and/or Li and euthanized. Urine and kidneys were collected for analysis. When administered with Li, CLPD ameliorated polyuria, attenuated the rise in urine prostaglandin E2 (PGE2), and resulted in significantly higher urinary arginine vasopressin (AVP) and aldosterone levels as compared to Li treatment alone. However, urine sodium excretion remained elevated. Semi-quantitative immunoblotting revealed that CLPD alone increased renal aquaporin 2 (AQP2), Na-K-2Cl cotransporter (NKCC2), Na-Cl cotransporter (NCC), and the subunits of the epithelial Na channel (ENaC) in medulla by 25–130 %. When combined with Li, CLPD prevented downregulation of AQP2, Na-K-ATPase, and NKCC2 but was less effective against downregulation of cortical α- or γ-ENaC (70 kDa band). Thus, CLPD primarily attenuated Li-induced downregulation of proteins involved in water conservation (AVP-sensitive), with modest effects on aldosterone-sensitive proteins potentially explaining sustained natriuresis. Confocal immunofluorescence microscopy revealed strong labeling for P2Y12-R in proximal tubule brush border and blood vessels in the cortex and less intense labeling in medullary thick ascending limb and the collecting ducts. Therefore, there is the potential for CLPD to be directly acting at the tubule sites to mediate these effects. In conclusion, P2Y12-R may represent a novel therapeutic target for Li-induced NDI.

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Acknowledgments

This work was supported by a grant from the US Department of Veterans Affairs Merit Review Program (to B. K. Kishore) and the resources and facilities at the VA SLC Health Care System, Salt Lake City, Utah, and Marriott Cardiovascular Fellowship (to C. M. Ecelbarger). Additional funding sources include National Institute of Diabetes and Digestive and Kidney Diseases Grant DK-64324 (to J. Peti-Peterdi) and an Established Investigator Award from the American Heart Association (to C. M. Ecelbarger).

Conflict of interest

No conflicts of interest, financial or otherwise, are declared by the author(s). Parts of this work were presented at the Kidney Week 2013 of the American Society of Nephrology, October–November 2013, Atlanta, GA, and appeared as a printed abstract in the proceedings of that meeting [49].

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    Authors and Affiliations

    1. Department of Internal Medicine and Center on Aging, University of Utah Health Sciences Center & Veterans Affairs Salt Lake City Health Care System, 500 Foothill Drive (151M), Salt Lake City, UT, 84148, USA

      Yue Zhang, Kristina M. Heiney & Bellamkonda K. Kishore

    2. Zilkha Neurogenetic Institute and Department of Physiology and Biophysics, University of Southern California, 1501 San Pablo Street, ZNI 313, Los Angeles, CA, 90033, USA

      János Peti-Peterdi & Anne Riquier-Brison

    3. PharmaCenter Bonn, Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, D-53121, Bonn, Germany

      Christa E. Müller

    4. Department of Neurobiology and Anatomy and Center on Aging Geriatric Research, Education, and Clinical Center (GRECC), University of Utah Health Sciences Center & Veterans Affairs Salt Lake City Health Care System, 500 Foothill Drive (151B), Salt Lake City, UT, 84148, USA

      Noel G. Carlson

    5. Department of Medicine, Center for the Study of Sex Differences in Health, Aging, and Disease, Georgetown University, 4000 Reservoir Road NW Bldg D, Rm 392, Washington, DC, 20057, USA

      Carolyn M. Ecelbarger

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    1. Yue Zhang
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    Correspondence to Bellamkonda K. Kishore.

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    Carolyn M. Ecelbarger and Bellamkonda K. Kishore contributed equally to this work.

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    Zhang, Y., Peti-Peterdi, J., Heiney, K.M. et al. Clopidogrel attenuates lithium-induced alterations in renal water and sodium channels/transporters in mice. Purinergic Signalling 11, 507–518 (2015). https://doi.org/10.1007/s11302-015-9469-0

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