解剖学报

解剖学报 ›› 2013, Vol. 44 ›› Issue (2 ): 163-169.doi: 10.3969/j.issn.0529-1356.2013.02.004

• 神经生物学 • 上一篇    下一篇

粒细胞集落刺激因子通过调节小胶质细胞活化改善缺氧致脑室周围白质损伤

杜阳 姚琳丽 杨阳 高明 郝爱军*    

  1. 山东大学医学院组织学胚胎学教研室，教育部实验畸形学重点实验室，济南250012
  • 收稿日期:2012-09-24 修回日期:2012-11-13 出版日期:2013-04-06 发布日期:2013-04-06
  • 通讯作者: 郝爱军 E-mail:aijunhao@sdu.edu.cn
  • 基金资助:

    国家自然科学基金重点资助项目;教育部高等学校博士学科点专项科研基金资助项目

Granulocyte-colony stimulating factor ameliorates periventricular white matter damage subject to hypoxia via influencing on activation of microglia

DU Yang YAO Lin-li YANG Yang GAO Ming HAO Ai-jun*    

  1. Key Laboratory of the Ministry of Education for Experimental Teratology, Department of Histology and Embryclogy, School of Medicine, Shandong University, Ji’nan 250012, China
  • Received:2012-09-24 Revised:2012-11-13 Online:2013-04-06 Published:2013-04-06

摘要：

目的 探讨粒细胞集落刺激因子通过影响小胶质细胞的活化对缺氧导致的脑室周围白质损伤（PWMD）的保护作用。方法 将1d龄新生小鼠108只随机分为对照组、损伤组及治疗组，后两组经缺氧箱缺氧法制备脑室周围白质损伤模型，造模前及造模后2h给予治疗组存活鼠腹腔注射粒细胞集落刺激因子，之后每日1次，1d、3d、7d后各处死3组部分动物。取全脑切片进行免疫荧光双标检测小胶质细胞的募集以及炎性因子的分泌情况；取脑室周围白质，采用酶联免疫吸附剂测定法检测炎性因子分泌水平；利用RT-PCR法检测致炎因子和抑炎因子的分泌以及两类活化小胶质细胞的数量和比例变化情况；治疗组结束给药于7d，分别于5d、8d、10d、12d、30d进行神经行为学实验，观察其感觉运动功能的发育。结果 粒细胞集落刺激因子能够促进小鼠运动功能恢复，改善脑瘫症状，改变活化小胶质细胞中M1型细胞和M2型细胞的数量和比例，使促炎性因子分泌降低，抑炎因子及神经营养因子分泌升高，改善损伤导致的神经发育异常及神经行为缺陷。结论 利用粒细胞集落刺激因子干预脑室周围白质损伤可以抗炎，并可以诱导小胶质细胞向神经保护方向转化，调节炎性因子和神经营养因子的分泌。

关键词: 缺氧 , 脑室周围白质损伤 , 粒细胞集落刺激因子 , 小胶质细胞 , 神经保护 , 免疫荧光 , 反转录-聚合酶链反应 , 小鼠

Abstract:

Objective To explore the protective effect of granulocyte colony stimulating factor (G-CSF) on the activation of microglial in periventricular white matter damage (PWMD) model mice. Methods One-day old newborn mice（n =108）were randomly divided into control group, injury group and treated group. PWMD model was produced by the hypoxia box in the injury group and treated group. Two hours later, the survived mice in the treated group accepted an intraperitoneal injection of rhG-CSF, continued for 7 days. Some mice in 3 groups were sacrificed on day 1, day 3 and day 7 after treatment. The whole brain of the rat was taken for histological analysis and immunofluorescence to observe the accumulation of microglia in vivo and the secretion of cytokines. The periventricular white matter was collected using enzyme-linked immunosorbent assay (ELISA) to detect the level of inflammatory cytokine secretion. The secretion of proinflammatory cytokines and suppression of inflammatory cytokines as well as two types of activated microglia in the number and proportion changes were detected by RT-PCR. The injections of rhG-CSF was finished on day 7 and the neurobehavioral experiments began on day 5, 8, 10, 12 and 30, respectively. The changes of their muscle tone, voluntary movement, and emotional behavior were observed. Results G-CSF facilitated the recruitment of microglia, changed the number and proportion of the M1 and M2 cells in activated microglia, and decreased proinflammatory cytokine secretion as well as increasing the neurotrophic factor secretion. In addition, the recovery of muscle tone, voluntary movement, and sense and motor fuctions were promoted by G-CSF. Conclusion In this study, the administration of G-CSF in the periventricular white matter injury accelerated mouse motor function recovery, improved cerebral palsy symptoms, led the microglia altering to neural protection, and regulated the secretion of inflammatory factors and neurotrophic factors.

Key words: Hypoxia , Periventricular white matterdamage , Granulocyte-colony stimulating factor , Microglia , Neural protection , Immunofluorescence , RT-PCR , Mouse