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

我们在NG-108 -15细胞中发现了2-甲酰胺醇是内皮素大麻素受体配体（CB 1受体），并在细胞中Cai-D12+甲酰胺D1集中的过渡酶，2-甲酰胺醇被添加到细胞暂停中。Twenty four structural analogs of 2-arachidonoylglyManagement ol were prepared and tested the ability of rise of CaイイD12+イエ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 D12+ y D1 in the cells began as low as 0.3 nM of the compound. Whereas HU-210和CP 55940，合成大麻素受体抑制剂，在0.3 nM的条件下提高了Cai-D12 + Cai-D1。The maximum rises of Ca D12 + y D1 were m\lower than that of 2-arachidonoylglycerol。上面的结果清楚地显示了2-甲酰胺醇是神经细胞中的完全拮抗剂和真正的内源性大麻素受体配体，下一步，我们研究了大脑中2-甲酰胺醇的生物合成途径。在大鼠脑同源性中，2-花生醇和二酰甘油醇含有花生酸在孵化过程中快速生成。当含有甲氧烷酸时，甲氧烷酸被添加到大鼠脑同源物中，2-甲氧烷醇被检测到。2-Arachidonoylglycerol是由放射性磷脂酰肌醇产生的，但不是由磷脂酰胆碱或磷脂酰亚胺产生的。这些数据强烈地提出了磷脂酰肌醇在磷脂酶C反应下水解至二酰甘油，并在二酰甘油脂酶中转化为2-甲氧烷醇。

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.