摘要
目的探讨脑出血(ICH)大鼠立体定向血肿抽吸术并行原位神经干细胞移植模型的建立方法及治疗作用。方法采用大鼠自体股动脉血(武汉大学人民医院动物实验中心)经自动注射仪注入尾状核的方法制作大鼠ICH模型。6 h后向血肿腔内注入尿激酶模拟临床微创血肿抽吸术清除血肿。90只SD大鼠随机数字表达分为5组,分别为假手术组(A组)、ICH模型组(B组)、ICH+血肿抽吸手术组(C组)、ICH+血肿抽吸+磷酸盐缓冲液(phosphate buffersaline,PBS)对照组(D组)、ICH+血肿抽吸+神经干细胞(NSCs)移植组(E组)。移植后3d干湿重法测量脑含水量。移植5d原位缺口末端标记法(TUNEL)检测大鼠血肿周边细胞凋亡水平。分别于3、7、14、28d对每组大鼠进行神经功能缺陷症状评分,14d后水迷宫实验检测其空间学习能力,对该模型及治疗效果进行评估,两组间样本均数比较采用t检验,多组间比较采用方差分析。结果成功建立ICH及血肿抽吸模型。脑含水量B组(83.49±1.49)高于E组(75.63±1.15,t=10.229,P<0.01),C组(79.66±1.64)高于E组(75.63±1.15,t=4.928,P<0.01)、血肿周围细胞凋亡水平B组(31.11±1.60)高于E组(12.01±1.06,t=24.376,P<0.05),C组(18.45±0.89)高于E组(12.01±1.06,t=11.397,P<0.05)、7天后神经功能评分A组(2.29±1.14)低于B组(12.25±2.56,t=8.706,P<0.01),C组(6.32±1.07)低于B组(12.25±2.56,t=5.235,P<0.01),E组(4.36±0.68)低于D组(5.35±0.48t=2.913,P<0.05)、水迷宫实验E组逃避潜伏期明显短于对照组(20.86±17.55,F=4.083,P<0.05),平台象限的游泳距离/总游泳距离的百分比低于对照组(26.48±11.25,F=7.955,P<0.01)。结论微创血肿抽吸联合神经干细胞移植对ICH有明显的协同神经功能保护作用。
Objective To investigate the establishment and therapeutic effect of stereotactic hemato-ma aspiration combined with orthotopic neural stem cell transplantation in rats with intracerebral hemorrhage(ICH).Methods The rat model of ICH was established by injecting autologous femoral artery blood into caudate nucleus by automatic injector.At 6 h later,urokinase was injected into the hematoma cavity to dis-solve blood clots,and minimally invasive hematoma aspiration was performed to remove the intracranial he-matoma.Totally,90 male Sprague-Dawley(SD)rats were randomly divided into five groups according to the random digits table(n=18 in each group):sham operation group(group A),ICH model group(group B),cerebral hemorrhage+hematoma aspiration group(group C),ICH+hematoma aspiration+normal phosphate buffer saline(PBS)group(group D),ICH+hematoma aspiration+neural stem cells(NSCs)transplantation group(group E).Brain water content was measured 3 days after transplantation.Cell apoptosis was examined by terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)on the 5th day after transplantation.The neurological defective scores were assessed at 3,7,14,28 days after ICH respectively in each experiment,and the spatial learning ability was detected by Morris Water Maze 14 days later.The model and the therapeutic effect were evaluated.The measurement data were expressed by mean±standard deviation,the sample mean size between the two groups was compared by t-test,and the sample mean between multiple groups was compared by one-way ANOVA.Results The models of ICH and hematoma aspiration were successfully established.The brain water content in group B(83.49±1.49)was higher than in group E(75.63±1.15,P<0.01),and that in group C(79.66±1.64)was higher than in group E(75.63±1.15,4.928,P<0.01).The apoptosis level around hematoma in group B(31.11±1.60)was higher than in group E(12.01±1.06,24.376,P<0.05),and that in group C(18.45±0.89)was higher than in group E(12.01±1.06,11.397,P<0.05).At 7th day,the neurological function score in group A(2.29±1.14)was lower than in group B(12.25±2.56,t=8.706,P<0.01),that in group C(6.32±1.07)was lower than in group B(12.25±2.56,t=5.235,P<0.01),and that in group E(4.36±0.68)was lower than in group D(5.35±0.48,t=2.913,P<0.05).MWM test showed that the escape latency in group E was significantly lower than that in other groups(20.86±17.55,F=4.083,P<0.05),and the percentage of swimming distance/total swimming distance in the platform quadrant in group E was lower than other groups(26.48±11.25,F=7.955,P<0.01).Conclusion Co-transplantation of NCSs and minimally invasive hematoma aspiration produces a synergistic neuroprotective effect for ICH.
作者
宋平
雷盼
蔡强
华秋伟
李知阳
魏航宇
周龙
高论
程利
罗明
陈谦学
Song Ping;Lei Pan;Cai Qiang;Hua Qiuwei;Li Zhiyang;Wei Hangyu;Zhou Long;Gao Lun;Cheng Li;Luo Ming;Chen Qianxue(Department of Neurosurgery,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Intensive Care Units,Renmin Hospital of Wuhan University,Wuhan 430060,China;Department of Neurosurgery,WuhanNo.1Hospital,Wuhan 430022,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第5期880-883,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81671306、81971158、82271518)
湖北省卫生健康科研基金(WJ2019H431)
武汉市卫生健康委员会科研项目(WX19B08)。
关键词
脑出血
立体定向血肿抽吸
神经干细胞
移植
行为学
Intracerebral hemorrhage
Stereotactic hematoma aspiration
Neural stem cells
Transplantation
Behavior