Cultured rat microglial cells synthesize the endocannabinoid 2-arachidonylglycerol, which increases proliferation via a CB2 receptor-dependent mechanism
Cultured rat microglial cells synthesize the endocannabinoid 2-arachidonylglycerol, which increases proliferation via a CB2 receptor-dependent mechanism
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DOI：
10.1124/mol.65.4.999
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发表时间：
2004-04-01
影响因子：
3.6
通讯作者：
Hillard, CJ
中科院分区：
文献类型：
--
作者：
Carrier, EJ;Kearn, CS;Hillard, CJ
作为中枢神经系统中的吞噬细胞和抗原呈递细胞，小胶质细胞在诸如中风和多发性硬化症的疾病过程中被激活。由于外周巨噬细胞能够产生内源性大麻素，我们已经研究了内源性大麻素的生产中的巨噬细胞集落刺激因子（M-CSF）依赖的大鼠小胶质细胞系（RTMGL 1）使用反相高压液相色谱和液相色谱-质谱。我们确定，培养的小胶质细胞产生的内源性大麻素2-花生四烯酸甘油（2-AG），以及花生四烯酸在较小的数量。当2-AG，而不是花生四烯酸，外源性添加，RTMGL 1小胶质细胞增加其增殖。这种增殖增加被CB 2受体拮抗剂N-[（1 S）内-1，3，3-三甲基双环庚-2-基]-5-（4-氯-3-甲基苯基）-1-（4-甲基苄基）-吡唑-3-甲酰胺（SR 144528）阻断，并被CB 2受体特异性激动剂1，1-二甲基丁基-1-脱氧-Delta（9）-四氢大麻酚（JWH 133）模拟。伴随2-AG增殖的增加，活性ERK 1也增加，这也被SR 144528阻断。RTMGL 1小胶质细胞，它存在于一个引发的状态，表达CB 1和CB 2受体所证明的逆转录-聚合酶链反应和免疫染色。未处理细胞中的CB 2受体在细胞表面和内部都表达，并且细胞暴露于2-AG显著增加受体内化。这些数据表明，CB 2受体的2-AG激活可能有助于小胶质细胞的增殖反应，如发生在神经退行性疾病。
Microglia, as phagocytes and antigen-presenting cells in the central nervous system, are activated in such disease processes as stroke and multiple sclerosis. Because peripheral macrophages are capable of producing endocannabinoids, we have examined endocannabinoid production in a macrophage-colony stimulating factor (M-CSF)-dependent rat microglial cell line (RTMGL1) using reversed phase high-pressure liquid chromatography and liquid chromatography-mass spectroscopy. We determined that cultured microglial cells produce the endocannabinoid 2-arachidonylglycerol (2-AG) as well as anandamide in smaller quantities. When 2-AG, but not anandamide, is added exogenously, RTMGL1 microglia increase their proliferation. This increased proliferation is blocked by an antagonist of the CB2 receptor N-[(1S)endo-1,3,3-trimethyl bicyclo heptan-2-yl]-5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)-pyrazole-3-carboxamide (SR144528) and mimicked by the CB2 receptor-specific agonist 1,1-dimethylbutyl-1-deoxy-Delta(9)-tetrahydrocannabinol (JWH133). Accompanying the increase in proliferation seen with 2-AG is an increase in active ERK1 that is also blocked with SR144528. The RTMGL1 microglial cells, which exist in a primed state, express the CB1 and CB2 receptors as demonstrated by reverse transcription-polymerase chain reaction and immunostaining. The CB2 receptor in untreated cells is expressed both at the cell surface and internally, and exposure of the cells to 2-AG significantly increases receptor internalization. These data suggest that 2-AG activation of CB2 receptors may contribute to the proliferative response of microglial cells, as occurs in neurodegenerative disorders.