重组人粒细胞集落刺激因子对脑出血大鼠脑水肿及血管再生的影响

Effects of recombinan human granulocyte colony-stimulating factor on brain edema and vascular regeneration in rats after intracerebral hemorrhage

背景:研究表明重组人粒细胞集落刺激因子可以减轻脑缺血后的脑水肿,并且可动员内皮前体细胞增加脑缺血区域新生血管生成,促进患者神经功能恢复。目的:探讨重组人粒细胞集落刺激因子对大鼠脑出血后脑水肿及新生血管的影响。设计、时间及地点:随机对照动物实验,于2006-03/11在泸州医学院中心实验室完成。材料:健康雄性SD大鼠144只,随机分为假手术组、脑出血组、治疗组,48只/组。重组人粒细胞集落刺激因子为深圳新鹏生物工程有限公司产品。方法:脑出血组与治疗组大鼠采用断尾取自体血方式建立脑出血模型,假手术组仅经注射点注入生理盐水。造模1h后,治疗组大鼠腹腔注射重组人粒细胞集落刺激因子60μg/kg。分别于干预后6h,12h,24h,48h,72h,7d时间点,每组各取8只大鼠,采用干湿重法测定大鼠组织脑含水量,采用SP、DAB显色法免疫组化检测CD34+血管的表达。主要观察指标:脑组织含水量动态变化,CD34+血管免疫组化结果。结果:与假手术组比较,脑出血组大鼠脑组织含水量显著升高(t=4.49,P<0.05),在48h,72h最明显;各时间点治疗组脑组织含水量均明显低于脑出血组(t=6.74,P<0.05)。与假手术组比较,脑出血组CD34+血管数明显增加(t=3.42,P<0.05)治疗组CD34+血管数明显多于脑出血组(t=6.07,P<0.05),且在72h,7d时差异尤为明显。结论:重组人粒细胞集落刺激因子作用于脑出血大鼠后,可减轻脑水肿程度,促进血肿周围新生血管生成。

BACKGROUND： Administration of recombinant human granulocyte colony-stimulating factor （rhG-CSF） is known to diminish cerebral edema and to enhance the new vascularization by mobilizing endothelial progenitor cells in cerebral infarction, then to promote the neurofunctional recovery. OBJECTIVE： To investigate the effects of rhG-CSF on brain edema and new vessels after intracerebral hemorrhage （ICH） in rats. DESIGN, TIME AND SETTING： The randomized controlled animal study was performed at the Central Laboratory, Luzhou Medical College from March to November 2006. MATERIALS： A total of 144 healthy male Sprague Dawley rats were equally and randomly assigned into a sham operation group a ICH group and a treatment group, rhG-CSF （Xinpeng, Shenzhen, China） was used in this study. METHODS： Rat models of ICH were made by the method of cutting off the tail of each rat to obtain autoblood in the ICH and treatment groups. Rats in the sham operation group were injected with saline. Rats in the treatment group were administered with rhG-CSF （60 μg/kg） by intraperitoneal injection after 1 hour. Eight rats from each group were studied at 6, 12, 24, 48, 72 hours and 7 days. Brain water content of rats was measured by dry-wet method. CD34^＋ vessel expression was detected by SP and DAB coloration, immunohistochemical method. MAIN OUTCOME MEASURES： Dynamic changes in brain water content; immunohistochemical results of CD34^＋ vessels. RESULTS： The water contents were significantly higher in the ICH group than in the sham operation group （t=4.49, P 〈 0.05）, reaching the peak at hours 48 and 72. The water contents were significantly lower in the treatment group than in the ICH group （t =6.74, P 〈 0.05）. The microvascular expressions of CD34 were higher in the ICH group than in the sham operation group （t=3.42, P 〈 0.05）. The microvascular expressions of CD34 were higher in the treatment group than in the ICH group （t=6.07, P 〈 0.05）, reaching the peak at 72 hours and 7 days. CONCLUSION： RhG-CSF could relieve brain edema and enhance new vessels generating in the peripheral tissues of hematoma region in ICH rats.