Yeon Soo-In, Kim Joo Young, Yeon Dong-Soo, Abramowitz Joel, Birnbaumer Lutz, Muallem Shmuel, Lee Young-Ho
Department of Physiology and BK21 PLUS Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Korea.
Department of Pharmacology and BK21 PLUS Project for Medical Sciences, Yonsei University College of Medicine, Seoul, Korea.
PLoS One. 2014 Oct 13;9(10):e110413. doi: 10.1371/journal.pone.0110413. eCollection 2014.
Transient receptor potential canonical type 3 (TRPC3) channels are non-selective cation channels and regulate intracellular Ca2+ concentration. We examined the role of TRPC3 channels in agonist-, membrane depolarization (high K+)-, and mechanical (pressure)-induced vasoconstriction and vasorelaxation in mouse mesenteric arteries. Vasoconstriction and vasorelaxation of endothelial cells intact mesenteric arteries were measured in TRPC3 wild-type (WT) and knockout (KO) mice. Calcium concentration ([Ca2+]) was measured in isolated arteries from TRPC3 WT and KO mice as well as in the mouse endothelial cell line bEnd.3. Nitric oxide (NO) production and nitrate/nitrite concentrations were also measured in TRPC3 WT and KO mice. Phenylephrine-induced vasoconstriction was reduced in TRPC3 KO mice when compared to that of WT mice, but neither high K+- nor pressure-induced vasoconstriction was altered in TRPC3 KO mice. Acetylcholine-induced vasorelaxation was inhibited in TRPC3 KO mice and by the selective TRPC3 blocker pyrazole-3. Acetylcholine blocked the phenylephrine-induced increase in Ca2+ ratio and then relaxation in TRPC3 WT mice but had little effect on those outcomes in KO mice. Acetylcholine evoked a Ca2+ increase in endothelial cells, which was inhibited by pyrazole-3. Acetylcholine induced increased NO release in TRPC3 WT mice, but not in KO mice. Acetylcholine also increased the nitrate/nitrite concentration in TRPC3 WT mice, but not in KO mice. The present study directly demonstrated that the TRPC3 channel is involved in agonist-induced vasoconstriction and plays important role in NO-mediated vasorelaxation of intact mesenteric arteries.
瞬时受体电位香草酸亚型3（TRPC3）通道是非选择性阳离子通道，可调节细胞内钙离子浓度。我们研究了TRPC3通道在激动剂、膜去极化（高钾）和机械（压力）诱导的小鼠肠系膜动脉血管收缩和舒张中的作用。在TRPC3野生型（WT）和基因敲除（KO）小鼠中测量了内皮细胞完整的肠系膜动脉的血管收缩和舒张。在TRPC3 WT和KO小鼠的分离动脉以及小鼠内皮细胞系bEnd.3中测量了钙离子浓度（[Ca2+]）。还在TRPC3 WT和KO小鼠中测量了一氧化氮（NO）生成以及硝酸盐/亚硝酸盐浓度。与WT小鼠相比，TRPC3 KO小鼠中去氧肾上腺素诱导的血管收缩减弱，但TRPC3 KO小鼠中高钾或压力诱导的血管收缩均未改变。TRPC3 KO小鼠以及选择性TRPC3阻滞剂吡唑-3抑制了乙酰胆碱诱导的血管舒张。在TRPC3 WT小鼠中，乙酰胆碱阻断了去氧肾上腺素诱导的钙离子比率升高及随后的舒张，但对KO小鼠的这些结果影响甚微。乙酰胆碱在内皮细胞中引起钙离子升高，这被吡唑-3抑制。乙酰胆碱在TRPC3 WT小鼠中诱导NO释放增加，但在KO小鼠中未增加。乙酰胆碱还使TRPC3 WT小鼠中的硝酸盐/亚硝酸盐浓度升高，但在KO小鼠中未升高。本研究直接证明，TRPC3通道参与激动剂诱导的血管收缩，并在完整肠系膜动脉的NO介导的血管舒张中起重要作用。
