大黄酚对原代培养小鼠星形胶质细胞谷氨酸代谢的影响

Effects of chrysophanol on glutamate metabolism of primarily cultured strocytes in mice

目的探讨不同浓度大黄酚对原代培养小鼠星形胶质细胞活性及谷氨酸代谢功能的影响。方法原代培养小鼠星形胶质细胞,大黄酚10、20、40、60、80、100、200、250 mg/L处理细胞1、2、3d,MTT法检测大黄酚对星形胶质细胞活性的影响;RT-PCR检测谷氨酸/天门冬氨酸转运体(GLAST)及谷氨酸转运体1(GLT-1)mRNA表达水平。相关试剂盒检测谷氨酰胺合成酶(GS)和谷胱甘肽过氧化物酶(GSH-Px)活性。结果大黄酚可抑制星形胶质细胞活性,可诱导星形胶质细胞GLAST和GLT-1 mRNA表达上调,GSH-Px和GS活性增加,且均具有时间和浓度依赖性。但相似浓度且作用时间相同的大黄酚并不增加细胞的活性,即大黄酚对细胞外谷氨酸浓度的影响并不依赖细胞数量或活性的增加。结论大黄酚可抑制小鼠星形胶质细胞活性,对谷氨酸代谢的调节具有时间和浓度依赖性。

Objective To study the effects of different concentration of chrysophanol on the activity and glutamate metabolism of primarily cultured astrocytes in vitro. Methods Primarily cultured mice astrocytes were treated with chrysophanol at concentration of 10,20,40,60,80,100,200,and 250 mg/L（ chry 1 ~ 8 group） for 1 d,2 d,and 3 d. The cell activities were measured by MTT method,mRNA expressions of glutamate/aspartate transporter（ GLAST） and glutamate acid transporter 1（ GLT-1） were detected by using RT-PCR,and the activities of glutamine synthetase（ GS） and glutathione peroxidase（ GSH-Px） were also determined. Results Time-dependence and concentration-dependence of the effects of chrysophanol showed on the following： activity of astrocytes inhibited,mRNA expressions of GLAST and GLT-1 upregulated,and the activities of GSH-Px and GS increased. However,the effect of chrysophanol on extra-cellular concentration of glutamate didn＇t depend on the number or activity of the cells. Conclusion Chrysophanol inhibited the activity of astrocytes in mice,and its effect on glutamate metabolism was concentration- and time-dependent.