ANANDAMIDE, AN ENDOGENOUS CANNABIMIMETIC EICOSANOID, BINDS TO THE CLONED HUMAN CANNABINOID RECEPTOR AND STIMULATES RECEPTOR-MEDIATED SIGNAL-TRANSDUCTION
ANANDAMIDE, AN ENDOGENOUS CANNABIMIMETIC EICOSANOID, BINDS TO THE CLONED HUMAN CANNABINOID RECEPTOR AND STIMULATES RECEPTOR-MEDIATED SIGNAL-TRANSDUCTION
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DOI：
10.1073/pnas.90.16.7656
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发表时间：
1993-08-15
影响因子：
11.1
通讯作者：
DEVANE, WA
中科院分区：
文献类型：
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作者：
FELDER, CC;BRILEY, EM;DEVANE, WA
花生四烯酸乙醇酰胺（anandamide），一个候选的内源性大麻素配体，最近已被分离出猪脑和显示大麻素样结合活性突触体膜制剂和模仿大麻素诱导的抑制抽搐反应在离体小鼠输精管。在这项研究中，大麻素和几种同源物被评估为大麻素激动剂，通过检查它们与克隆的大麻素受体结合的能力，抑制毛喉素刺激的cAMP积累，抑制N型钙通道，并刺激一种或多种功能性第二信使反应。合成的大麻素和除一种同类物外的所有同类物竞争[H-3] CP 55，940与从表达大鼠大麻素受体的L细胞制备的质膜结合。在表达人大麻素受体（HCR，称为CHO-HCR细胞）的中国仓鼠卵巢（CHO）细胞中评价大麻素激活受体介导的信号转导的能力，并与表达毒蕈碱m5受体的对照CHO细胞（CHOm 5细胞）进行比较。Anandamide抑制毛喉素刺激的cAMP在CHO-HCR细胞中的积累，但在CHOm 5细胞中不抑制，并且这种反应被百日咳毒素阻断。花生四烯酸酰胺和几种活性同类物在N18神经母细胞瘤细胞中抑制N型钙通道。花生四烯酸和细胞内钙的释放，花生四烯酸刺激CHOm 5和CHO-HCR细胞，并没有影响释放肌醇磷酸或磷脂酰乙醇，产生后激活磷脂酶C和D，分别。大麻素似乎表现出被归类为大麻素/大麻素受体激动剂所需的基本标准，并与其他大麻素激动剂一样对花生四烯酸和细胞内钙释放具有相似的非受体效应。
Arachidonylethanolamide (anandamide), a candidate endogenous cannabinoid ligand, has recently been isolated from porcine brain and displayed cannabinoid-like binding activity to synaptosomal membrane preparations and mimicked cannabinoid-induced inhibition of the twitch response in isolated murine vas deferens. In this study, anandamide and several congeners were evaluated as cannabinoid agonists by examining their ability to bind to the cloned cannabinoid receptor, inhibit forskolin-stimulated cAMP accumulation, inhibit N-type calcium channels, and stimulate one or more functional second messenger responses. Synthetic anandamide, and all but one congener, competed for [H-3]CP55,940 binding to plasma membranes prepared from L cells expressing the rat cannabinoid receptor. The ability of anandamide to activate receptor-mediated signal transduction was evaluated in Chinese hamster ovary (CHO) cells expressing the human cannabinoid receptor (HCR, termed CHO-HCR cells) and compared to control CHO cells expressing the muscarinic m5 receptor (CHOm5 cells). Anandamide inhibited forskolin-stimulated cAMP accumulation in CHO-HCR cells, but not in CHOm5 cells, and this response was blocked with pertussis toxin. N-type calcium channels were inhibited by anandamide and several active congeners in N18 neuroblastoma cells. Anandamide stimulated arachidonic acid and intracellular calcium release in both CHOm5 and CHO-HCR cells and had no effect on the release of inositol phosphates or phosphatidylethanol, generated after activation of phospholipase C and D, respectively. Anandamide appears to exhibit the essential criteria required to be classified as a cannabinoid/anandamide receptor agonist and shares similar nonreceptor effects on arachidonic acid and intracellular calcium release as other cannabinoid agonists.