Gupta Saurabh, Lozano-Cuenca Jair, Villalón Carlos M, de Vries René, Garrelds Ingrid M, Avezaat Cees J J, van Kats Jorge P, Saxena Pramod R, MaassenVanDenBrink Antoinette
Department of Pharmacology, Erasmus MC, University Medical Center Rotterdam, 3000 DR, Rotterdam, The Netherlands.
Naunyn Schmiedebergs Arch Pharmacol. 2007 Mar;375(1):29-38. doi: 10.1007/s00210-007-0137-y. Epub 2007 Feb 13.
Capsaicin, a pungent constituent from red chilli peppers, activates sensory nerve fibres via transient receptor potential vanilloid receptors type 1 (TRPV1) to release neuropeptides like calcitonin gene-related peptide (CGRP) and substance P. Capsaicin-sensitive nerves are widely distributed in human and porcine vasculature. In this study, we examined the mechanism of capsaicin-induced relaxations, with special emphasis on the role of CGRP, using various pharmacological tools. Segments of human and porcine proximal and distal coronary arteries, as well as cranial arteries, were mounted in organ baths. Concentration response curves to capsaicin were constructed in the absence or presence of the CGRP receptor antagonist olcegepant (BIBN4096BS, 1 microM), the neurokinin NK1 receptor antagonist L-733060 (0.5 microM), the voltage-sensitive calcium channel blocker ruthenium red (100 microM), the TRPV1 receptor antagonist capsazepine (5 microM), the nitric oxide synthetase inhibitor Nomega-nitro-L-arginine methyl ester HCl (L-NAME; 100 microM), the gap junction blocker 18alpha-glycyrrhetinic acid (10 microM), as well as the RhoA kinase inhibitor Y-27632 (1 microM). Further, we also used the K+ channel inhibitors 4-aminopyridine (1 mM), charybdotoxin (0.5 microM) + apamin (0.1 microM) and iberiotoxin (0.5 microM) + apamin (0.1 microM). The role of the endothelium was assessed by endothelial denudation in distal coronary artery segments. In distal coronary artery segments, we also measured levels of cyclic adenosine monophosphate (cAMP) after exposure to capsaicin, and in human segments, we also assessed the amount of CGRP released in the organ bath fluid after exposure to capsaicin. Capsaicin evoked concentration-dependent relaxant responses in precontracted arteries, but none of the above-mentioned inhibitors did affect these relaxations. There was no increase in the cAMP levels after exposure to capsaicin, unlike after (exogenously administered) alpha-CGRP. Interestingly, there were significant increases in CGRP levels after exposure to vehicle (ethanol) as well as capsaicin, although this did not induce relaxant responses. In conclusion, the capsaicin-induced relaxations of the human and porcine distal coronary arteries are not mediated by CGRP, NK1, NO, vanilloid receptors, voltage-sensitive calcium channels, K+ channels or cAMP-mediated mechanisms. Therefore, these relaxant responses to capsaicin are likely to be attributed to a non-specific, CGRP-independent mechanism.
辣椒素是红辣椒中的一种辛辣成分,它通过瞬时受体电位香草酸受体1(TRPV1)激活感觉神经纤维,从而释放降钙素基因相关肽(CGRP)和P物质等神经肽。辣椒素敏感神经广泛分布于人体和猪的血管系统中。在本研究中,我们使用各种药理学工具,研究了辣椒素诱导血管舒张的机制,特别关注了CGRP的作用。将人体和猪的近端及远端冠状动脉节段以及颅动脉节段安装在器官浴槽中。在不存在或存在CGRP受体拮抗剂olcegepant(BIBN4096BS,1微摩尔)、神经激肽NK1受体拮抗剂L-733060(0.5微摩尔)、电压敏感性钙通道阻滞剂钌红(100微摩尔)、TRPV1受体拮抗剂辣椒平(5微摩尔)、一氧化氮合酶抑制剂盐酸Nω-硝基-L-精氨酸甲酯(L-NAME;100微摩尔)、缝隙连接阻滞剂18α-甘草次酸(10微摩尔)以及RhoA激酶抑制剂Y-27632(1微摩尔)的情况下构建辣椒素的浓度-反应曲线。此外,我们还使用了钾通道抑制剂4-氨基吡啶(1毫摩尔)、蝎毒素(0.5微摩尔)+蜂毒明肽(0.1微摩尔)以及iberiotoxin(0.5微摩尔)+蜂毒明肽(0.1微摩尔)。通过剥脱远端冠状动脉节段的内皮来评估内皮的作用。在远端冠状动脉节段中,我们还测量了暴露于辣椒素后环磷酸腺苷(cAMP)的水平,在人体节段中,我们还评估了暴露于辣椒素后器官浴液中释放的CGRP量。辣椒素在预收缩的动脉中引起浓度依赖性的舒张反应,但上述抑制剂均未影响这些舒张反应。与(外源性给予的)α-CGRP不同,暴露于辣椒素后cAMP水平没有升高。有趣的是,暴露于溶剂(乙醇)以及辣椒素后CGRP水平均显著升高,尽管这并未诱导舒张反应。总之,辣椒素诱导的人体和猪远端冠状动脉舒张并非由CGRP、NK1、一氧化氮、香草酸受体、电压敏感性钙通道、钾通道或cAMP介导的机制所介导。因此,这些对辣椒素的舒张反应可能归因于一种非特异性的、不依赖CGRP的机制。