大麻素对星形胶质细胞谷氨酸释放的影响及机制研究  

Effect of Cannabinoids HU210 on Glutamate Release of Astrocytes and Its Mechanism

作　　者：徐逸[1] 韩静[2] 王伟[3] 朱舟[3] 

机构地区：[1]华中科技大学同济医学院附属同济医院整形外科,武汉430030 [2]陕西师范大学现代教学技术教育部重点实验室,西安710062 [3]华中科技大学同济医学院附属同济医院神经内科,武汉430030

出　　处：《神经损伤与功能重建》2017年第1期1-4,共4页Neural Injury and Functional Reconstruction

基　　金：湖北省科技支撑计划(No.015BKA220);华中科技大学自主创新研究基金(No.2015ZHYX011)

摘　　要：目的:观察人工合成大麻素HU210对体外培养中脑腹侧被盖(VTA)区星形胶质细胞谷氨酸释放的影响,并探讨抑制星形胶质细胞谷氨酸释放的药物治疗途径。方法:体外培养大鼠VTA脑区星形胶质细胞,RT-PCR及免疫荧光染色检测胶质纤维酸性蛋白(GFAP)及大麻素受体1(CB1R)的表达。将培养的星形胶质细胞分为4组:对照组(培养基中只加入0.2%DMSO),HU210组(培养基中加入3μM HU210),HU210+AM281组(培养基同时加入3μM HU210及3μM AM281)和HU210+Riluzole组(培养基同时加入3μM HU210及3μM Riluzole),各组4孔。干预30 min后,采用谷氨酸检测试剂盒观察各组星形胶质细胞培养基中谷氨酸浓度。再培养VTA脑区星形胶质细胞,分为对照组(培养基中只加入0.2%DMSO)和Riluzole组(培养基中加入3μM Riluzole),干预30 min后,利用Western Blot印迹分析Riluzole干预后谷氨酸转运体-1(GLT-1)表达水平的变化。结果:RT-PCR及免疫荧光染色显示,星形胶质细胞内有广泛CB1R的表达。与对照组比较,HU210组星形胶质细胞谷氨酸的释放显著增加(P<0.01),HU210+AM281组及HU210+Riluzol组星形胶质细胞谷氨酸的释放较HU210组均显著降低(P<0.01);且HU210+Riluzol组星形胶质细胞的GLT-1表达较对照组升高(P<0.05)。结论:HU210可能通过激活星形胶质细胞的CB1R促进星形胶质细胞释放谷氨酸。Riluzole可能通过升高星形胶质细胞的GLT-1的表达,逆转HU210所致的谷氨酸释放。Objective： To investigate the effect of HU210 on glutamate release of cultured midbrain ventral tegmental area （VTA） astrocytes and its mechanism. Methods： VTA astrocytes were cultured. RT-PCR and immunostaining were used to detect the expression of glia fibrillary acidic protein （GFAP） and cannabinoid 1 receptor （CB1R）. The cultured VTA astroeytes were randomly divided into 4 groups （n=4）： control group （0.2%DMSO in medium）, HU210 group （3 μM HU210 in medium）, HU210＋AM281group （3 μM HU210 ＋ 3 μM AM281 in medium） and HU210＋Riluzole group （3 μM HU210 ＋ 3 μM Riluzole in medium）. Concentration of glutamate in medium was detected using enzyme immunoassay of glutamate kit. And then the cultured VTA astrocytes were randomly divided into 2 groups （n=3）： control group （0.2%DMSO in medium） and Riluzole group （3 μM Riluzole in medium）. Western Bolt was used to detect the expression of glutamate transporter-1 （GLT-1）. Results： Abundant CB1R RNA and the protein were detected in astrocytes. HU210 increased glutamate release from cultured VTA astrocytes, which was suppressed by AM281. Riluzole suppressed HU210-induced glutamate release from cultured VTA astrocytes and increased the expression of astrocytic GLT-1 which served to remove the released glutamate. Conclusion： Cannabinoids HU210 probably activates VTA astrocytic CB1R to release glutamate and Riluzole probably increases the expression of GLT-1, thus suppressing HU210-induced glutamate release.

关 键 词：星形胶质细胞 HU210 谷氨酸 大麻素受体1 利鲁唑 

分 类 号：R741[医药卫生—神经病学与精神病学] R971[医药卫生—临床医学]