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Endothelial cell Orai1 is essential for endothelium-dependent contraction of mouse carotid arteries in normotensive and hypertensive mice

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Abstract

Endothelium-dependent contraction (EDC) exists in blood vessels of normotensive animals, but is exaggerated in hypertension. An early signal in EDC is cytosolic Ca2+ rise in endothelial cells. In this study we investigated the functional role of Orai1, a major endothelial cell Ca2+ entry channel, in EDC. Hypertension model was established in WT mice by intake of L-NNA in the drinking water (0.5 g/L) for 4 weeks or osmotic pump delivery of Ang II (1.5 mg·kg−1·d−1) for 2 weeks. In TRPC5 KO mice, the concentration of L-NNA and Ang II were increased to 1 g/L or 2 mg·kg−1·d−1, respectively. Arterial segments were prepared from carotid arteries and aortas, and EDC was elicited by acetylcholine in the presence of Nω-nitro-L-arginine methyl ester. We showed that low concentration of acetylcholine (3–30 nM) initiated relaxation in phenylephrine-precontracted carotid arteries of both normotensive and hypertensive mice, while high concentration of acetylcholine (0.1–2 μM) induced contraction. Application of selective Orai1 inhibitors AnCoA4 (100 μM) or YM58483 (400 nM) had no effect on ACh-induced relaxation but markedly reduced acetylcholine-induced EDC. We found that EDC was increased in hypertensive mice compared with that of normotensive mice, which was associated with increased Orai1 expression in endothelial cells of hypertensive mice. Compared to TRPC5 and TRPV4, which were also involved in EDC, endothelial cell Orai1 had relatively greater contribution to EDC than either TRPC5 or TRPV4 alone. We identified COX-2, followed by PGF2α, PGD2 and PGE2 as the downstream signals of Orai1/TRPC5/TRPV4. In conclusion, Orai1 coordinates together with TRPC5 and TRPV4 in endothelial cells to regulate EDC responses. This study demonstrates a novel function of Orai1 in EDC in both normotensive and hypertensive mice, thus providing a general scheme about the control of EDC by Ca2+-permeable channels.

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Fig. 1: Orai1 participates in ACh-induced EDC in carotid arteries of normotensive mice.
Fig. 2: Lack of effect for Orai1 inhibitors on TP receptor-mediated smooth muscle contraction in carotid arteries of normotensive mouse.
Fig. 3: Orai1 participates in ACh-induced EDC in carotid arteries of L-NNA-induced hypertensive mice.
Fig. 4: Orai1 contributes to ACh-induced Ca2+ entry in primary endothelial cells of mouse carotid arteries.
Fig. 5: TRPC5 and TRPV4 participate in ACh-induced EDC in carotid arteries of normotensive mice.
Fig. 6: TRPC5 and TRPV4 participate in ACh-induced EDC in carotid arteries of L-NNA-induced hypertensive mice.
Fig. 7: Essential role of COX-2 in the EDC of carotid arteries in normotensive and hypertensive mice.
Fig. 8: Orai1, TRPC5 and TRPV4 contribute to the production of multiple prostanoids in carotid arteries of L-NNA-induced hypertensive mice.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This work was supported by grants from Hong Kong Research Grant Committee (14100619, RIF/R4005-18F) and Innovation and Technology Fund ITS/212/21.

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Authors and Affiliations

  1. School of Biomedical Sciences, Heart and Vascular Institute and Li Ka Shing Institute of Health Science, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China

    Xiao Li, Zhen-chuan Lei, Chun Yin Lo, Tsz Yau Jan, Chi Wai Lau & Xiao-qiang Yao

  2. Centre for Cell & Developmental Biology, School of Life Sciences, The Chinese University of Hong Kong, Hong Kong, China

    Xiao-qiang Yao

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  1. Xiao Li
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Contributions

XQY revised the manuscript, supervised the study, and obtained funding; XL contributed to the experiment design, conducted the experiments, performed data analyzes, prepared the figures, and drafted the manuscript. ZCL, CYL, TYJ, and CWL conducted the experiments. All authors contributed to the final manuscript and approved submission.

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Correspondence to Xiao-qiang Yao.

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The authors declare no competing interests.

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All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals issued by the US National Institute of Health (NIH) and approved by the Animal Experimentation Ethics Committee, Chinese University of Hong Kong.

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Li, X., Lei, Zc., Lo, C.Y. et al. Endothelial cell Orai1 is essential for endothelium-dependent contraction of mouse carotid arteries in normotensive and hypertensive mice. Acta Pharmacol Sin 45, 975–987 (2024). https://doi.org/10.1038/s41401-024-01227-6

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