粒细胞集落刺激因子对急性脊髓损伤中小胶质细胞的作用  

Effect of Granulocyte-colony stimulating factor on microglial function in the acute spinal cord injury mice model

作　　者：张慧[1] 郭雨霁[1] 郝爱军[1] 马保华[1] 

机构地区：[1]山东大学医学院组织胚胎学教研室,济南250012

出　　处：《山东大学学报（医学版）》2012年第5期10-14,19,共6页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金资助项目(Y2004C08)

摘　　要：目的探讨粒细胞集落刺激因子对急性脊髓损伤条件下小胶质细胞神经保护/损伤功能转变的影响。方法建立小鼠半切脊髓损伤模型。体内实验是造模成功术后第1天开始给予粒细胞集落刺激因子,连续3 d,进行Basso-Beattie-Bresnahan(BBB)评分后处死,取脊髓切片进行免疫荧光检测小胶质细胞的募集。体外实验是取小鼠脊髓的组织上清液,加到小胶质细胞株BV-2培养基中共培养24 h,治疗组加入粒细胞集落刺激因子12 h后收集细胞。采用RT-PCR和免疫印迹法,检测小胶质细胞炎症因子的分泌及信号通路的改变。结果粒细胞集落刺激因子能够促进急性脊髓损伤后运动功能的恢复,促进小胶质细胞募集。体外实验结果显示,粒细胞集落刺激因子可以降低小胶质细胞NF-κB信号通路p65的活性,改变小胶质细胞因子的分泌,促炎性因子IL-1β表达降低,抗炎因子TGFβ表达升高。结论应用粒细胞集落刺激因子治疗脊髓损伤可以促进小鼠运动功能的恢复并调节小胶质细胞的分布和功能,诱导小胶质细胞向神经保护的方向转化。Objective To explore the alteration of microglial protection/injury function caused by granulocyte-colony stimulating factor（G-CSF） in the acute spinal cord injury mice model.Methods Hemi-section injuries were produced by cutting one-half of the right lateral portion of the spinal cord.After surgery the mice were given an intraperitoneal injection of rhG-CSF,which continued 3 days.The Basso-Beattie-Bresnahan（BBB） locomotor rating scale was used to analyze behavior and then the spinal cord was taken out for histological analysis by immunofluorescence to observe the accumulation of microglia in vivo.Microglial cell line BV-2 cells were pretreated with normal and injury spinal cord supernatant for 24 h,followed by a 12 h administration of G-CSF（100 ng/mL）,then harvested the cells.RT-PCR and Western blot were employed to evaluate the changes of microglial cytokines and pathways in vitro.Results In the G-CSF treatment group,improvement in hind-limb motor function and accumulation of microglial cell were significantly greater than in the sham-treatment（vehicle） group.As the results in vitro showed,in the G-CSF treatment group,the activity of NF-κB p65 was inhibited and the secretion of microglial cytokines was changed.In addition,expression of pro-inflammation cytokine IL-1β decreased and anti-inflammation cytokine TGFβ increased.Conclusions In this study,the administration of G-CSF in acute spinal cord injury not only improved motor function,but also regulated the distribution and function of microglia as well as to induce the microglia altering to neural protection.

关 键 词：脊髓损伤 粒细胞集落刺激因子 小神经胶质细胞 神经保护药 

分 类 号：R329.3[医药卫生—人体解剖和组织胚胎学]