Study on the 2-arachidonoylglycerol, a novel endogenous cannabinoid receptor ligand

新型内源性大麻素受体配体2-花生四烯酰甘油的研究

基本信息

批准号：
10470488
负责人：
WAKU Keizo
金额：
$ 3.07万
依托单位：
Teikyo University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10470488/
关键词：
cannabinoid arachidonoylglycerol monoglyceride anadamide arachidonic acid

项目摘要

We have found that 2-arachidonoylglycerol is the endogenous cannabinoid receptor ligand (CB1 receptor) in NG-108-15 cells and caused the transient increase in CaィイD12+ィエD1 concentration in the cells when 2-arachidonoylglycerol was added to the cell suspension. Twenty four structural analogs of 2-arachidonoylglycerol were prepared and tested the ability of rise of CaィイD12+ィエD1 in the cells. It was demonstrated that 2-arachidonoylglycerol was found to be the most potent compound after testing all of the compounds and the rise of CaィイD12+ィエD1 in the cells began as low as 0.3 nM of the compound. Whereas HU-210 and CP55940, the synthetic cannabinoid receptor agonists, raised the CaィイD12+ィエD1 at 0.3 nM,. The maximum rises of CaィイD12+ィエD1 were much lower than that of 2-arachidonoylglycerol. The above results clearly showed that 2-arachidonoylglycerol is the full agonist and real endogenous cannabinoid receptor ligand in the neuronal cells.Next, we investigated the biosynthetic route of 2-arachidonoylglycerol in the brain. In the rat brain homogenate, 2-arachidonoylglycerol and diaylglycerol containing arachidonic acid was rapidly generated during incubation. When diaylglycerol containing arachidonic acid was added to the rat brain homogenate, 2-arachidonoylglycerol was detected. 2-Arachidonoylglycerol was produced from radioactive phosphatidylinositol, but not from phosphatidylcholine or phosphatidylethanolamine. These data strongly suggest that phosphatidylinositol was hydrolyzed to diacylglycerol by phospholipase C action and converted into 2-arachidonoylglycerol by diacylgllycerol lipase.

我们已经发现，NG-108-15细胞中的2-芳基二烯丙基甘油是内源性大麻素受体配体（CB1受体），当将2-芳基二烯酰基甘油醇添加到细胞中时，导致细胞中CAIY D12+IE D1浓度的短暂增加。制备了24个2-芳基酮甘油的结构类似物，并测试了细胞中Caiy D12+IE D1的升高能力。事实证明，在测试所有化合物后，发现2-芳基二烯丙基甘油是最潜在的化合物，并且细胞中CAI D12+IE D1的上升开始低至该化合物的0.3 nm。而HU-210和CP55940（合成大麻素受体激动剂）将CAI D12+IE D1提高到0.3 nm，。 CAI D12+IE D1的最大升高远低于2-芳基烯丙基甘油。以上结果清楚地表明，2-芳基烯丙基甘油是神经元细胞中的全部激动剂和真实的内源性大麻素受体配体。我们研究了大脑中2-芳基烯丙基甘油的生物合成途径。在大鼠脑匀浆中，在孵育过程中迅速产生含有花生四烯酸的2-芳烃。当将含有蛛网膜酸的二酰甘油醇添加到大鼠脑匀浆中时，检测到2-芳基烯丙基甘油。由放射性磷脂酰胆碱或磷脂酰乙醇胺产生的2-芳基烯丙基甘油。这些数据强烈表明，通过磷脂酶C作用将磷脂酰甘油醇水解为二酰基甘油，并通过二酰基甘油脂肪酶转化为2-芳基酮甘油。