Department of Psychological and Brain Sciences, Indiana University, Bloomington,IN 47405, USA.
Br J Pharmacol. 2012 Apr;165(8):2414-24. doi: 10.1111/j.1476-5381.2011.01497.x.
Endometriosis is a disorder in which the endometrium forms growths outside the uterus and is associated with chronic pain. Recent evidence suggests that endometrial motility plays a role in the aetiology of endometriosis. The endocannabinoid system regulates cellular migration. Given the growing involvement of the endocannabinoids in reproduction, we investigated the role of the endocannabinoid system in migration of endometrial cells.
Migration of the human endometrial HEC-1B cells was assayed. Standard PCR techniques were used to determine the presence of the GPCR, GPR18, in HEC-1B cells, and p44/42 MAPK was assayed in stably transfected HEK293-GPR18 cells to determine receptor specificity for known cannabinoid agonists and antagonists. N-arachidonoyl ethanolamine (AEA) metabolism was measured, using HPLC/MS/MS for lipid analysis.
AEA, Δ(9) -tetrahydrocannabinol (Δ(9) -THC) and N-arachidonoyl glycine (NAGly) induce migration of HEC-1B cells through cannabinoid CB(1) receptor-independent mechanisms. MAPK activation in HEK293-GPR18 cells revealed novel pharmacology for known CB(1) and CB(2) receptor ligands at GPR18 receptors, including Δ(9) -THC, which activates MAPK at nanomolar concentrations, whereas WIN 55212-2, CP55940, JWH-133 and JWH-015, and arachidonyl-1-hydroxy-2-propylamide (R1-methanandamide) had no effect. Moreover, HEC-1B migration and MAPK activation by NAGly and Δ(9) -THC were antagonized by Pertussis toxin, AM251 and cannabidiol.
An understanding of the function and regulation of GPR18 and its molecular interactions with endogenous ligands, and how phytocannabinoids play a role with GPR18 signalling is vital if we are to comprehensively assess the function of the cannabinoid signalling system in human health and disease.
This article is commented on by Alexander, pp. 2411-2413 of this issue and is part of a themed section on Cannabinoids in Biology and Medicine. To view Alexander visit http://dx.doi.org/10.1111/j.1476-5381.2011.01731.x. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2012.165.issue-8. To view Part I of Cannabinoids in Biology and Medicine visit http://dx.doi.org/10.1111/bph.2011.163.issue-7.
子宫内膜异位症是一种子宫内膜在子宫外生长的疾病，与慢性疼痛有关。最近的证据表明，子宫内膜的运动在子宫内膜异位症的发病机制中起作用。内源性大麻素系统调节细胞迁移。鉴于内源性大麻素在生殖中的作用不断增加，我们研究了内源性大麻素系统在子宫内膜细胞迁移中的作用。
检测人子宫内膜 HEC-1B 细胞的迁移。使用标准 PCR 技术确定 HEC-1B 细胞中 GPCR、GPR18 的存在，并在稳定转染的 HEK293-GPR18 细胞中检测 p44/42 MAPK，以确定已知大麻素激动剂和拮抗剂对受体的特异性。使用 HPLC/MS/MS 进行脂质分析来测量 N-花生四烯酰乙醇胺 (AEA) 的代谢。
AEA、Δ(9)-四氢大麻酚 (Δ(9)-THC) 和 N-花生四烯酰甘氨酸 (NAGly) 通过大麻素 CB1 受体非依赖性机制诱导 HEC-1B 细胞迁移。在 HEK293-GPR18 细胞中 MAPK 的激活揭示了已知 CB1 和 CB2 受体配体在 GPR18 受体上的新药理学，包括 Δ(9)-THC，其以纳摩尔浓度激活 MAPK，而 WIN 55212-2、CP55940、JWH-133 和 JWH-015 以及花生四烯酸-1-羟基-2-丙酰胺 (R1-甲氨酰胺) 则没有影响。此外，NAGly 和 Δ(9)-THC 诱导的 HEC-1B 迁移和 MAPK 激活被百日咳毒素、AM251 和大麻二酚拮抗。
如果我们要全面评估大麻素信号系统在人类健康和疾病中的功能和调节，以及植物大麻素如何与 GPR18 信号发挥作用，那么了解 GPR18 的功能和调节及其与内源性配体的分子相互作用是至关重要的。
本文由 Alexander 在本期 2411-2413 页进行了评论，并且是关于大麻素在生物学和医学中的主题部分的一部分。要查看 Alexander，请访问 http://dx.doi.org/10.1111/j.1476-5381.2011.01731.x。要查看该部分中的其他文章，请访问 http://dx.doi.org/10.1111/bph.2012.165.issue-8。要查看大麻素在生物学和医学中的第 I 部分，请访问 http://dx.doi.org/10.1111/bph.2011.163.issue-7。
