摘要
目的 研究人重组粒细胞集落刺激因子（rG-CSF）对骨髓干细胞（BMSCs）在脑缺血大鼠血液和脑组织中的分布变化及抗脑缺血损伤的影响.方法 将106只SD大鼠按随机数字表法分为假手术组（10只）、模型组（48只）、rG-CSF组（48只）,后两组再分为术后2、3、7、14 d亚组,每个亚组12只.用改良线栓法制备大鼠局灶性脑缺血模型.rG-CSF组于术前3 d和术后2 d皮下注射rG-CSF 10μg/kg,假手术组和模型组给予等量生理盐水,均每日1次.术后各时间点进行神经功能评分;取腹主动脉血,测定外周血白细胞计数（WBC）及CD34＋细胞计数;观察脑组织病理改变;并用免疫组化法测定脑组织CD34＋细胞表达.结果 ①制模后2 d大鼠神经功能评分即显著降低,随后逐渐升高;rG-CSF组术后7 d和14 d神经功能评分（分）较模型组显著增高（7 d：11.86±0.69比10.53±0.76,14 d：13.38±0.52比12.38±0.52,均P〈0.01）.②制模后2 dSb周血WBC和CD34＋细胞计数即显著增加,3 d达峰值,7 d和14 d降低;除14 d CD34＋细胞计数外,rG-CSF组其余时间点WBC和CD34＋细胞计数均较模型组明显增加[WBC（×109/L）2 d：11.75±1.76比8.07±1.27,3 d：13.07±1.70比10.88±1.78,7 d：8.63±1.36比5.58±1.57,14 d：6.98±0.98比4.87±0.92;CD34＋细胞计数（个/μl）2 d：8.83±2.14比3.17±0.75,3 d：13.50±1.87比5.00±1.55,7 d：5.33±1.21比2.33±1.21,P〈0.05或P〈0.013.③制模后2 d大鼠脑组织CD34＋细胞表达即明显增强,7 d达峰值,14 d降低;rG-CSF组各时间点CD34＋表达[吸光度（A）值]均较模型组显著增加（2 d：43.21±4.41比22.04±2.95,3 d：45.79±1.76比25.69±2.44,7 d：52.09±2.86比33.04±2.62,14 d：29.73±1.99比16.91±2.95,均P〈0.01）.④rG-CSF组脑组织病理损伤较模型组减轻,以14 d改善明显.结论 脑缺血可引起BMSCs进入外周血及向脑组织归巢,其在外周血和脑组织的变化呈先增多再减少的特点,分别于缺血后3 d和7 d达峰值;rG-CSF可使进入外周血和脑组织的BMSCs明显增加.BMSCs动员对脑缺血损伤的保护作用明显,且随动员后时间的延长呈增强趋势.
Objective To explore the influence of recombination granulocyte colony stimulating factor （rG-CSF）on mobilization and distribution of bone marrow stem cells （BMSCs） in blood and brain tissue,and its role in protecting brain in rats with cerebral ischemia.Methods One hundred and six SpragueDawley（SD）rats were divided into sham-operated group （n=10）,model group（n=48）,rG-CSF group （n=48） according to the method of random digital table,and rats in model and rG-CSF groups were divided into four subgroups：i.e.2,3,7 and 14 days subgroups,with 12 rats in each subgroup.Middle cerebral artery occlusion（MCAO）model was reproduced with nylon thread.In rats of rG-CSF group rG-CSF （10 btg/kg）was administered by subcutaneous injection 3 days before and 2 days after operation respectively,once a day.Rats in sham-operated and model groups were administered with normal saline in the same volume,once a day.At the corresponding time after operation,general neural function score（GNFS）of rats was measured.Blood was collected through abdominal aorta,then white blood cell （WBC） and CD34＋ cells in peripheral blood were counted.Brain pathologic changes were observed,and expression of CD34＋ cells in rats brain tissue was determined by using immunohistochemical method.Results ①GNFS was lower obviously in 2-day model group compared with that in sham-operated group,and then increased gradually.At 7 days and 14 days after operation,GNFS in rG-CSF group was higher significantly than that in model group （7 days：11.86±0.69 vs.10.53±0.76,14 days：13.38±0.52 vs.12.38±0.52,both P〈0.01）.②WBC and CD34＋ cells in peripheral blood in model group increased obviously,with the highest level appeared at 3 days and lowered at 7 days and 14 days.Increase of WBC and CD34＋ cells in rats of rG-CSF group was more obvious than that of model group at each time point except CD34＋ in 14 days group [WBC （×109/L）2 days：11.75±1.76 vs.8.07±1.27,3 days：13.07±1.70 vs.10.88±1.78,7 days：8.63±1.36 vs.5.58士1.57,14 days：6.98士0.98 vs.4.87士0.92;CD34＇（cells/t^1）2 days：8.83±2.14 vs.3.17±0.75,3 days：13.50±1.87 vs.5.00±1.55,7 days：5.33±1.21 vs.2.33±1.21,P〈0.05 or P〈0.01].③Expression of CD34＋ cells in the brain of rats in 2-day model group increased significantly,and the highest level appeared at 7 days and decreased at 14 days.Absorbance （A） value of CD34＋ cells expression in rat brains of each rG-CSF group was more significant than that in model group（2 days：43.21±4.41 vs.22.04±2.95,3 days：45.79±1.76 vs.25.69±2.44,7 days：52.09±2.86 vs.33.04±2.62,14 days：29.73±1.99 vs.16.91±2.95,all P〈0.01）.④ The signs of injury to brain in pathological examination were less obvious in 14 days rG-CSF group.Conclusion BMSCs could be induced to enter peripheral blood and ＂home＂ to brain tissue after cerebral ischemia.It was showed that BMSCs increased in number at first and then decreased in peripheral blood and brain,the peak number was found on 3rd day in peripheral blood and 7th day in brain.Mobilization with rG-CSF could increase the number of BMSCs in peripheral blood and brain tissue.The effect of mobilization of BMSCs on protecting brain was significant after cerebral ischemia,and effect appeared to be more pronounced with prolongation of mobilization.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2011年第6期333-336,共4页
Chinese Critical Care Medicine
基金
国家自然科学基金资助项目（30973744）
河南省高校新世纪优秀人才支持计划项目（2006HANCET-05）
关键词
脑缺血
粒细胞集落刺激因子
骨髓干细胞
CD34+
Cerebral ischemia
Granulocyte colony stimulating factor
Bone marrow stem cell
CD34＋
