内源性大麻素在缺乏窖蛋白-1 的小胶质细胞系中脂质筏的区室化。
Compartmentalization of endocannabinoids into lipid rafts in a microglial cell line devoid of caveolin-1.
机构信息
Department of Neurobiology, Weizmann Institute of Science, Rehovot, Israel.
出版信息
Br J Pharmacol. 2012 Apr;165(8):2436-49. doi: 10.1111/j.1476-5381.2011.01380.x.
BACKGROUND AND PURPOSE
N-acyl ethanolamines (NAEs) and 2-arachidonoyl glycerol (2-AG) are endogenous cannabinoids and along with related lipids are synthesized on demand from membrane phospholipids. Here, we have studied the compartmentalization of NAEs and 2-AG into lipid raft fractions isolated from the caveolin-1-lacking microglial cell line BV-2, following vehicle or cannabidiol (CBD) treatment. Results were compared with those from the caveolin-1-positive F-11 cell line.
EXPERIMENTAL APPROACH
BV-2 cells were incubated with CBD or vehicle. Cells were fractionated using a detergent-free continuous OptiPrep density gradient. Lipids in fractions were quantified using HPLC/MS/MS. Proteins were measured using Western blot.
KEY RESULTS
BV-2 cells were devoid of caveolin-1. Lipid rafts were isolated from BV-2 cells as confirmed by co-localization with flotillin-1 and sphingomyelin. Small amounts of cannabinoid CB(1) receptors were found in lipid raft fractions. After incubation with CBD, levels and distribution in lipid rafts of 2-AG, N-arachidonoyl ethanolamine (AEA), and N-oleoyl ethanolamine (OEA) were not changed. Conversely, the levels of the saturated N-stearoyl ethanolamine (SEA) and N-palmitoyl ethanolamine (PEA) were elevated in lipid raft fractions. In whole cells with growth medium, CBD treatment increased AEA and OEA time-dependently, while levels of 2-AG, PEA and SEA did not change.
CONCLUSIONS AND IMPLICATIONS
Whereas levels of 2-AG were not affected by CBD treatment, the distribution and levels of NAEs showed significant changes. Among the NAEs, the degree of acyl chain saturation predicted the compartmentalization after CBD treatment suggesting a shift in cell signalling activity.
LINKED ARTICLES
This article is part of a themed section on Cannabinoids in Biology and Medicine. 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.
背景与目的
N-酰基乙醇胺（NAEs）和 2-花生四烯酰基甘油（2-AG）是内源性大麻素，与相关脂质一样，可按需从膜磷脂合成。在此，我们研究了缺乏窖蛋白-1 的小胶质细胞系 BV-2 中，大麻二酚（CBD）处理前后 NAEs 和 2-AG 向脂筏区室的定位。并将结果与窖蛋白-1 阳性的 F-11 细胞系进行了比较。
实验方法
BV-2 细胞用 CBD 或载体孵育。使用无去污剂连续 OptiPrep 密度梯度分离细胞。使用 HPLC/MS/MS 定量各馏分中的脂质。使用 Western blot 测定蛋白质。
主要结果
BV-2 细胞缺乏窖蛋白-1。通过与小窝蛋白-1 和神经酰胺共定位，证实 BV-2 细胞中分离出了脂筏。在脂筏中发现了少量大麻素 CB1 受体。孵育 CBD 后，2-AG、N-花生四烯酰乙醇胺（AEA）和 N-油酰乙醇胺（OEA）在脂筏中的水平和分布没有改变。相反，饱和 N-硬脂酰乙醇胺（SEA）和 N-棕榈酰乙醇胺（PEA）的水平在脂筏中升高。在含生长培养基的全细胞中，CBD 处理使 AEA 和 OEA 随时间依赖性增加，而 2-AG、PEA 和 SEA 的水平没有改变。
结论和意义
虽然 CBD 处理未影响 2-AG 的水平，但 NAEs 的分布和水平显示出显著变化。在 NAEs 中，酰基链饱和度的程度预示着 CBD 处理后的区室化，表明细胞信号转导活性发生了变化。
相关文章
本文是“大麻素在生物学和医学中的作用”主题部分的一部分。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作用”的第一部分，请访问 http://dx.doi.org/10.1111/bph.2011.163.issue-7。
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