1] Departamento de Bioquímica y Biología Molecular, Instituto Universitario de Investigación en Neuroquímica (IUIN), Facultad de Medicina, Universidad Complutense, Madrid, Spain [2] Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas (CIBERNED), Madrid, Spain [3] Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Madrid, Spain.
Cell Death Dis. 2013 Oct 17;4(10):e862. doi: 10.1038/cddis.2013.387.
The cannabinoid CB2 receptor, which is activated by the endocannabinoid 2-arachidonoyl-glycerol (2-AG), protects striatal neurons from apoptotic death caused by the local administration of malonate, a rat model of Huntington's disease (HD). In the present study, we investigated whether endocannabinoids provide tonic neuroprotection in this HD model, by examining the effect of O-3841, an inhibitor of diacylglycerol lipases, the enzymes that catalyse 2-AG biosynthesis, and JZL184 or OMDM169, two inhibitors of 2-AG inactivation by monoacylglycerol lipase (MAGL). The inhibitors were injected in rats with the striatum lesioned with malonate, and several biochemical and morphological parameters were measured in this brain area. Similar experiments were also conducted in vitro in cultured M-213 cells, which have the phenotypic characteristics of striatal neurons. O-3841 produced a significant reduction in the striatal levels of 2-AG in animals lesioned with malonate. However, surprisingly, the inhibitor attenuated malonate-induced GABA and BDNF deficiencies and the reduction in Nissl staining, as well as the increase in GFAP immunostaining. In contrast, JZL184 exacerbated malonate-induced striatal damage. Cyclooxygenase-2 (COX-2) was induced in the striatum 24 h after the lesion simultaneously with other pro-inflammatory responses. The COX-2-derived 2-AG metabolite, prostaglandin E2 glyceryl ester (PGE2-G), exacerbated neurotoxicity, and this effect was antagonized by the blockade of PGE2-G action with AGN220675. In M-213 cells exposed to malonate, in which COX-2 was also upregulated, JZL184 worsened neurotoxicity, and this effect was attenuated by the COX-2 inhibitor celecoxib or AGN220675. OMDM169 also worsened neurotoxicity and produced measurable levels of PGE2-G. In conclusion, the inhibition of 2-AG biosynthesis is neuroprotective in rats lesioned with malonate, possibly through the counteraction of the formation of pro-neuroinflammatory PGE2-G, formed from COX-2-mediated oxygenation of 2-AG. Accordingly, MAGL inhibition or the administration of PGE2-G aggravates the malonate toxicity.
大麻素 CB2 受体可被内源性大麻素 2-花生四烯酰甘油(2-AG)激活,可保护纹状体神经元免于局部施用丙二酸盐(一种亨廷顿病(HD)的大鼠模型)引起的细胞凋亡。在本研究中,我们通过检查二酰基甘油脂肪酶(催化 2-AG 生物合成的酶)的抑制剂 O-3841 以及 2-AG 失活的抑制剂 JZL184 或 OMDM169 的作用,研究了内源性大麻素是否在这种 HD 模型中提供紧张性神经保护, 2-AG 通过单酰基甘油脂肪酶(MAGL)。在丙二酸盐损伤纹状体的大鼠中注射抑制剂,并在该脑区测量几种生化和形态参数。在体外培养的具有纹状体神经元表型特征的 M-213 细胞中也进行了类似的实验。O-3841 可显著降低丙二酸盐损伤动物纹状体中的 2-AG 水平。但是,令人惊讶的是,抑制剂减轻了丙二酸盐引起的 GABA 和 BDNF 缺乏以及尼氏染色减少,并增加了 GFAP 免疫染色。相比之下,JZL184 加剧了丙二酸盐引起的纹状体损伤。伤后 24 小时,纹状体中同时诱导环加氧酶-2(COX-2)和其他促炎反应。COX-2 衍生的 2-AG 代谢物,前列腺素 E2 甘油酯(PGE2-G)加剧了神经毒性,而用 AGN220675 阻断 PGE2-G 作用可拮抗该作用。在暴露于丙二酸盐的 M-213 细胞中,COX-2 也被上调,JZL184 加重了神经毒性,而 COX-2 抑制剂塞来昔布或 AGN220675 可减轻该作用。OMDM169 也加重了神经毒性并产生可测量水平的 PGE2-G。总之,丙二酸盐损伤的大鼠中 2-AG 生物合成的抑制具有神经保护作用,可能是通过拮抗 COX-2 介导的 2-AG 氧化形成的促神经炎症性 PGE2-G 来实现的。因此,MAGL 抑制或 PGE2-G 的给药会加重丙二酸盐的毒性。