不同参数重复磁刺激对小鼠星形胶质细胞内源性大麻素相关基因表达的影响

Effects of different stimulus parameters of repeatitive magnetic stimulation on expression of ndocannabinoid- related gene in cultured mice astrocytes

目的探讨不同刺激参数重复磁刺激对小鼠星形胶质细胞内源性大麻素相关基因内源性大麻素受体l（Cannabinoid Receptor1，CBlR）、二酰基甘油脂肪酶（Diacylglycerol，DAGL）、脂肪酰胺水解酶（Fatty Acid Amide Hydrolase，FAAH）和单酰基甘油脂肪酶（Monoacylgiycerol，MAGL）表达的影响。方法从新生子鼠大脑中分离星形胶质细胞，待7～10d细胞长满进行纯化后，给予细胞不同参数的重复磁刺激或对照处理。磁刺激组分为4组，分别给予细胞频率为1Hz或5Hz，刺激强度为0．86T或1．24T的磁刺激处理，即（1Hz＋0．86T，1Hz＋1．24T，5Hz＋0．86T）和（5Hz＋1．24T），对照组细胞置于距重复磁刺激组细胞20cm的相同环境中，但是检测不到磁场强度。干预结束后，从细胞中提取总mRNA和蛋白，检测CB1R、DAGL、FAAH和MAGL的表达变化。结果与对照组相比，当给予细胞（5Hz＋0．86T）磁刺激处理时，小鼠星形胶质细胞的CBIR、MAGL和FAAH基因表达水平显著上调；当给予细胞（5Hz＋1．24T）磁刺激处理时，小鼠星形胶质细胞的FAAH基因表达水平显著上调；此外，4组刺激参数的重复磁刺激处理对小鼠星形胶质细胞的DAGL基因表达水平无显著影响。结论不同参数的重复磁刺激对小鼠星形胶质细胞内源性大麻素相关基因CB1R、DAGL、FAAH和MAGL基因表达水平的影响不同，不同参数磁刺激所致生物学效应差异可能与此有关。

Objective To explore the effects of different stimulus parameters of repetitive magnetic stimulation on the expression of endocannabinoid system-related genes, including eannabinoid receptor 1 （CBIR）, diacylglycerol （DAGL）, fatty acid amide hydrolase （FAAH） and monoaeylglyeerol （MAGL） in cultured mice astrocytes. Methods The astroeytes were isolated from the brain of newbom mice. After 7-10 d culture, the ceils were purified and then treated with repeatitive magnetic stimulation with different stimulus parameters or control treatment. The magnetic stimulation group was divided into four groups： （1 Hz ＋ 0.86T, 1 Hz ＋ 1.24 T, 5 Hz ＋ 0.86 T） and （5 Hz ＋ 1.24 T）, which represented the stimulation including the magnetic frequency of 1 Hz or 5 Hz and the intensity of 0.86 or 1.24 Tesla. The control cells were placed in the same environment but 20 cm away from the magnetic field of the repeated magnetic stimulation group, and the magnetic field strength could not be detected. After the intervention, total mRNA and protein were extracted from the cells and the expression changes of CB1R, DAGL, FAAH and MAGL were detected. Results Compared with the control group, the expression levels of CB 1R, MAGL and FAAH genes of astrocytes were significantly up-regulated when the ceils were stimulated with （5 Hz ＋ 0.86 T） magnetic stimulation. Meanwhile, the expression of FAAH gene were also up-regulated when the cells was stimulated with magnetic stimulation at （5 Hz ＋ 1.24 T）. However, there was no significant difference on the expression of DAGL between each group. Conclusions The effects of repeatitive magnetic stimulation of different parameters repeatitive magnetic stimulation induced different influence on the expression of endoeannabinoid system-related gene in astroeytes, which might be involved in its different therapeutic effects induced by different parameters.