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Antagonism of Ca2+-sensing receptors by NPS 2143 is transiently masked by p38 activation in mouse brain bEND.3 endothelial cells

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Abstract

Ca2+-sensing receptors (CaSR) are G protein-coupled receptors which are activated by a rise in extracellular Ca2+. CaSR activation has been known to inhibit parathyroid hormone release and stimulate calcitonin release from parathyroid glands and thyroid parafollicular C cells, respectively. The roles of CaSR in other cell types including endothelial cells (EC) are much less understood. In this work, we demonstrated protein and functional expression of CaSR in mouse cerebral EC (bEND.3). Unexpectedly, CaSR response (high Ca2+-elicited cytosolic [Ca2+] elevation) was unaffected by edelfosine or U73122 but strongly suppressed by SK&F 96365, ruthenium red, and 2-aminoethoxydiphenyl borate (2-APB), suggesting involvement of TRPV and TRPC channels but not Gq-phospholipase C. Acute application of NPS2143, a negative allosteric modulator of CaSR, suppressed CaSR response. However, a 40-min NPS2143 pre-treatment surprisingly enhanced CaSR response. After 4–24 h of application, this enhancement faded away and suppression of CaSR response was observed again. Similar results were obtained when La3+ and Sr2+ were used as CaSR agonists. The transient NPS 2143 enhancement effect was abolished by SB203580, a p38 inhibitor. Consistently, NPS 2143 triggered a transient p38 activation. Taken together, results suggest that in bEND.3 cells, NPS 2143 caused acute suppression of CaSR response, but then elicited a transient enhancement of CaSR response in a p38-dependent manner. NPS 2143 effects on CaSR in bEND.3 cells therefore depended on drug exposure time. These findings warrant cautious use of this agent as a CaSR modulator and potential cardiovascular drug.

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Acknowledgments

Y.M.L., K.L.W, and K.S.C. would like to thank China Medical University, China Medical University Hospital, Taiwan, and the Ministry of Science and Technology of Taiwan for providing funding (103-2320-B-039-015-; 104-2320-B-039-030-; 104-2320-B-039-013-; 105-2320-B-039-028-; 106-2320-B-039-025-; DMR-106-086; DMR-106-089; DMR-107-197; DMR-107-083; CMU107-S-01).

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  1. Ka-Shun Cheng, Paul Chan, and Yuk-Man Leung contributed equally as corresponding authors.

Authors and Affiliations

  1. School of Pharmacy, China Medical University, Taichung, 40402, Taiwan

    Cing-Yu Chen, Mann-Jen Hour & Wen-Chuan Lin

  2. Department of Anesthesiology, China Medical University Hospital, Taichung, Taiwan

    Kar-Lok Wong & Ka-Shun Cheng

  3. Department of Physiology, China Medical University, Taichung, 40402, Taiwan

    Lian-Ru Shiao & Yuk-Man Leung

  4. Department of Anesthesiology, The Qingdao University Yuhuangding Hospital, Yantai, Shandong, China

    Ka-Shun Cheng

  5. Division of Cardiology, Department of Medicine, Taipei Medical University Wan Fang Hospital, Taipei, Taiwan

    Paul Chan

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  1. Cing-Yu Chen
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Contributions

CYC and LRS conducted the experiments. WCL, KLW, MJH, and KSC participated in research design and data analysis. PC, CYC, and YML wrote the paper. All authors read and approved the manuscript.

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Correspondence to Yuk-Man Leung.

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No humans or animals were used in this study; only cell lines were used in this work and therefore ethical approval is not required.

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Chen, CY., Hour, MJ., Lin, WC. et al. Antagonism of Ca2+-sensing receptors by NPS 2143 is transiently masked by p38 activation in mouse brain bEND.3 endothelial cells. Naunyn-Schmiedeberg's Arch Pharmacol 392, 823–832 (2019). https://doi.org/10.1007/s00210-019-01637-y

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