马桑内酯对在体和离体小胶质细胞CD11b/c表达的影响

EFFECT OF CORIARIA LACTONE ON CD11b/c EXPRESSION OF MICROGLIA IN VIVO AND IN VITRO

目的研究致痫剂马桑内酯(CL)对在体和离体小胶质细胞CD11b/c表达的影响。方法①正常SD大鼠行马桑内酯侧脑室注射,观察大鼠的行为改变;利用免疫荧光染色的方法观察大鼠大脑皮质、海马内CD11b/c表达的变化。②纯化培养的小胶质细胞无血清培养,给予马桑内酯(5×10-5mol/L)刺激,利用免疫荧光染色结合流式细胞仪检测CD11b/c的表达。结果①马桑内酯侧脑室注射30min后均出现强烈的癫痫样发作,持续约4h;②马桑内酯侧脑室注射后大脑皮质及海马各区CD11b/c阳性细胞表达均出现明显增强,4-6h为表达高峰,至24h大脑皮质恢复至正常水平,但海马各区仍保持较高水平。③纯化培养的小胶质细胞在马桑内酯作用1h出现CD11b/c表达增强,2h达到高峰,至24h恢复正常。结论马桑内酯对小胶质细胞具有直接的活化作用;小胶质细胞的活化参与了马桑内酯的致痫过程。

Objective To explore the effect of convulsive agent Coriaria Lactone （CL） on the ex pression of CD11 b/c of microglia in vivo and in vitro. Methods ①CL was injected into the lateral ventricle of SD rats and the behaviour changes were observed. Immunofluorescent staining was used to assess the changes of CD11b/c expression in the cerebral cortex and hippocampus. ②Purified microglidwere cultured in serum-free condition and were treated with CL （5×10^-5mol/L）. Immunofluorescent staining and flow cytometry were used to asses the changes of CD11b/c expression. Results ① Injection of CL into animals resulted in robust behavioral seizures lasting between 30min and 4h. ②The expression of CD11b/c was significant enhanced in the cerebral cortex and hippocampus after CL injection. It appeared at lh, peaked at 4 or 6h, and gradually returned to normal in the cortex at 24h, but maintained a high level in the hippocampus. QThe expression of CD11b/c was significantly enhanced in purified microglia in vitro after CL treatment. It appeared at 1h, peaked at 2h and returned to normal at 24h. Conclusion The epileptogenic effect of CL involves a direct activation of microglia by the convulsive agent.