粒细胞集落刺激因子对慢性脑缺血大鼠神经功能的影响  

Effects of granulocyte colony-stimulating factor on brain nerve in chronic cerebral ischemic rats

作　　者：孔朝红[1] 刘煜敏[1] 朱江[1] 周绍霞[1] 李芹[1] 

机构地区：[1]武汉大学中南医院神经内科,430070

出　　处：《中华老年医学杂志》2013年第8期882-885,共4页Chinese Journal of Geriatrics

基　　金：国家自然科学基金（30470601）

摘　　要：目的观察粒细胞集落刺激因子（G-CSF）对慢性脑缺血大鼠行为学的影响，并探讨其可能的机制。方法制备大鼠慢性脑缺血双侧颈动脉结扎（2-VO）模型，随机分为假手术对照组和手术组，将手术组大鼠随机分为粒细胞集落刺激因子（G-CSF）组和磷酸盐缓冲液（PBS）组。术后6周G-CSF组经颈外静脉注射G-CSF（10mg／L，1ml·kg-1·d-1），PBS组给予PBS（1ml／kg），每间隔24h干预1次，共3次，假手术对照组仅分离出双侧颈外静脉，不干预。术后8周行Morris水迷宫评价大鼠空间学习记忆功能变化，同时通过观察缺血区细胞增生情况、激光共聚焦三维血管成像、缺血区神经细胞的凋亡和形态学变化以及血管内皮生长因子（VEGF）浓度等探讨其可能的机制。结果水迷宫检测结果显示，训练第2天至第5天大鼠逃避潜伏期G-CSF组明显短于PBS组（均P〈0．05）；第1象限游泳时间G-CSF组明显长于PBS组（P〈O．05）。缺血脑组织内的BrdU阳性细胞数G-CSF组（27．7±4．76）个／视野，明显高于PBS对照组（10．4±3．7）个／视野（P=0．030）。脑血管共聚焦检测结果显示，G-CSF组与PBS组比较，毛细血管直径明显变小（（2．90±0．20）μm与（3．45±0．26）μm，P=0．0203，同源组织缺血边界地区的分支点数目显著增加[（207．82±10．73）个／0．002mm2与（162．10士9。31）个／0．002mm3，P=0．0053，微血管总面积显著增加[（86498±2896）Ftm2／0．002ITlm。与（73976±3826）μm3／0．002mm3，P=0．003]。凋亡检测结果显示，G-CSF组细胞凋亡数目（32．10±6．70）个／视野，较PBS组（56．30±11．20）个／视野明显减少（F=11．89，P=0．043）。电镜下形态学观察结果显示，G-CSF组大鼠的细胞间隙炎性水肿明显减轻；G—CSF组大鼠血浆VEGF水平（58．81±6．61）ng／L，较PBS组（20．81±4．35）ng／L增加（P=0．025）。结论G-CSF可显著改�Objective To explore the effects of granulocyte colony stimulating factor （G-CSF） on chronic cerebral ischemia in rats, and its possible mechanism. Methods Chronic cerebral ischemia （2-VO） model was prepared and bilateral external jugular veins were isolated. A total of 30 rats were divided into 2 groups at random sham group （received no intervention, n= 15） and operative group （received G-CSF or PBS through external jugular vein injeetion, n= 15）. At 6 weeks after operation, the rats in operative group were divided into G-CSF group （received G-CSF 10 mg/L, 1 ml· kg 1 . d 1, 1 times every 24 h for, 3 times） and PBS control group （received PBS 10 mg/L,1 ml·kg 1 . d 11, 1 times every 24 h for 3 times）. At 8 weeks after the operation, morris water maze was carried out to evaluate the learning and memory ability of the rats. The cell proliferation, three- dimensional vascular distribution, ischemic neuronal apoptosis, cell morphological changes in ischemic area and the plasma VEGF levels were detected to explore the possible mechanisms. Results In morris water maze, escape latency at the 2rd to 5th day were significantly lower in G-CSF group than the PBS group （all P〈0.05）. The swimming time spent in the first quadrant in C^CSF group was significantly longer than the PBS group （P〈0.05）. There was a significant difference in the number of BrdU positive cells in the ischemical area between the G-CSF group and the control group [（27.7± 4.76） vs. （ 10.4± 3.7）, P = 0. 030） . Three-dimensional quantitative measurements of vascular structure showed that the capillary diameters was smaller in the G-CSF group than in the PBS group （（2.90± 0.20） μm vs. （3.45 ± 0.26） μm, P〈 0. 020] and the number of branch points in the boundary regions of ischemia had a significant difference in the CCSF group compared with the control group [（207.82±10.73） /0.002 mm3 vs. （162.10±9.31） /0.002mm3, P=0.005]. Three- dimensional cerebral vessel surface area in the ipsilate

关 键 词：粒细胞集落刺激因子 脑缺血 血管内皮生长因子类 

分 类 号：R743[医药卫生—神经病学与精神病学]