粒细胞集落刺激因子对小鼠急性脊髓损伤的神经保护作用及其机制  

Neuronal protective effect of granulocyte-colony stimulating factor and its related mechanism in acute spinal cord injury

作　　者：张慧[1] 郭雨霁[1] 邴鲁军[1] 刘尚明[1] 郝爱军[1] 

机构地区：[1]山东大学医学院组织胚胎学教研室,实验畸形学教育部重点实验室,济南250012

出　　处：《解剖学报》2012年第6期734-738,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金(青年)资助项目(81100919);山东省优秀科学家奖励基金资助项目(BS2010Y041)

摘　　要：目的探讨粒细胞集落刺激因子(G-CSF)对急性脊髓损伤条件下神经保护作用的具体机制。方法建立小鼠半切脊髓损伤模型。体内实验于造模成功术后第1天开始,给予G-CSF,连续3天,进行Basso-Beattie-Bresnahan(BBB)评分后处死,取脊髓组织进行免疫荧光及免疫印迹(Western blotting)检测。体外培养神经元,建立机械损伤细胞模型,进行形态学观察及缺口末端标记(TUNEL)染色。结果 G-CSF能够促进急性脊髓损伤后运动功能的修复,粒细胞集落刺激因子受体与核磷蛋白(NPM1)特异性表达在神经元上,而且G-CSF的使用能促进NPM1的表达,损伤脊髓组织内的凋亡神经元减少。体外实验结果显示,使用核磷蛋白特异性的抑制剂NSC348884能降低G-CSF对神经元的保护作用,凋亡细胞增多。结论在急性脊髓损伤中应用G-CSF可以促进损伤脊髓运动功能的修复,对神经元有明显的保护作用,这种保护作用可能是通过NPM1的抗凋亡作用而实现的。Objective To explore the mechanism of granulocyte-colony stimulating factor （G-CSF） on neuronal protective function in acute spinal cord model mice. Methods Hemi-section injury was produced by cutting one-half of the right lateral portions of the spinal cord. After the surgery the mouse accepted an intraperitoneal injection of recombinant human granulocyte colony-stimulating factor（rhGCSF） , once a day for 3 days. Behavior of the mouse was analyzed with Basso-Beattie-Bresnahan（BBB） locomotor rating scale. The spinal cord of the mouse was then dissected and examined with histological immunofluoreseence and Western blotting analysis. We established neuron mechanical injury model in vitro. Morphological ehanges were observed and TUNEL staining was used to investigate the apoptosis. Results In the G-CSF group, improvement in hindlimb motor funetion was significantly greater than that in the injury group. G-CSF receptor was expressed specifically on the neurons. After treatment with G-CSF in acute spinal cord injury, nucleophosmin 1 （NPMI） expression was significantly increased and apoptotic cells decreased. After treatment with NSC348884, a specific nucleophosmin inhibitor, in neuron mechanical injury model in vitro, apoptotic neurons increased and the function of G-CSF on neuronal protection was decreased. Conclusion G-CSF improves motor function in acute spinal cord injury. Moreover, this protective effect may be achieved through the anti-apoptotic role of NPM 1.

关 键 词：脊髓损伤 粒细胞集落刺激因子 核磷蛋白1 缺口末端标记染色 小鼠 

分 类 号：R651.2[医药卫生—外科学]