Smart D, Jerman J C, Gunthorpe M J, Brough S J, Ranson J, Cairns W, Hayes P D, Randall A D, Davis J B
Neuroscience Research, SmithKline Beecham Pharmaceuticals, New Frontiers Science Park, Third Avenue, Essex CM19 5AW, Harlow, UK.
Eur J Pharmacol. 2001 Apr 6;417(1-2):51-8. doi: 10.1016/s0014-2999(01)00901-3.
A full pharmacological characterisation of the recently cloned human vanilloid VR1 receptor was undertaken. In whole-cell patch clamp studies, capsaicin (10 microM) elicited a slowly activating/deactivating inward current in human embryonic kidney (HEK293) cells stably expressing human vanilloid VR1 receptor, which exhibited pronounced outward rectification (reversal potential -2.1+/-0.2 mV) and was abolished by capsazepine (10 microM). In FLIPR-based Ca(2+) imaging studies the rank order of potency was resiniferatoxin>olvanil>capsaicin>anandamide, and all were full agonists. Isovelleral and scutigeral were inactive (1 nM-30 microM). The potencies of capsaicin, olvanil and resiniferatoxin, but not anandamide, were enhanced 2- to 7-fold at pH 6.4. Capsazepine, isovelleral and ruthenium red inhibited the capsaicin (100 nM)-induced Ca(2+) response (pK(B)=6.58+/-0.02, 5.33+/-0.03 and 7.64+/-0.03, respectively). In conclusion, the recombinant human vanilloid VR1 receptor stably expressed in HEK293 cells acted as a ligand-gated, Ca(2+)-permeable channel with similar agonist and antagonist pharmacology to rat vanilloid VR1 receptor, although there were some subtle differences.
对最近克隆的人类香草酸受体1（VR1）进行了全面的药理学特性研究。在全细胞膜片钳研究中，辣椒素（10微摩尔）在稳定表达人类香草酸受体1的人胚肾（HEK293）细胞中引发了一种缓慢激活/失活的内向电流，该电流表现出明显的外向整流（反转电位为-2.1±0.2毫伏），并被辣椒平（10微摩尔）消除。在基于荧光成像板读数器（FLIPR）的钙离子成像研究中，效力顺序为：树脂毒素>奥伐尼>辣椒素>花生四烯酸乙醇胺，且均为完全激动剂。异维勒醇和黄贝母碱无活性（1纳摩尔至30微摩尔）。在pH 6.4时，辣椒素、奥伐尼和树脂毒素（而非花生四烯酸乙醇胺）的效力增强了2至7倍。辣椒平、异维勒醇和钌红抑制了辣椒素（100纳摩尔）诱导的钙离子反应（pK(B)分别为6.58±0.02、5.33±0.03和7.64±0.03）。总之，在HEK293细胞中稳定表达的重组人类香草酸受体1作为一种配体门控的、钙离子可通透的通道，其激动剂和拮抗剂药理学与大鼠香草酸受体1相似，尽管存在一些细微差异。
