神经干细胞治疗新生小鼠缺血缺氧性脑病损伤模型的行为学观察

摘要

目的 观测新生小鼠缺血缺氧性脑病(hypoxic-ischemic encephalopathy,HIE)经神经干细胞(neural stem cells,NSC)治疗后的行为学变化,为NSC治疗HIE提供行为学支持,并验证神经行为学实验Rotarod Test、Cylinder Test和Morris水迷宫对神经功能损伤恢复检测的适用性.方法 30只新生7d小鼠按随机数字表法分为3组.HIE+NSC组(治疗组):10只,先制成HIE模型,然后给予NSC治疗；HIE+ PBS组(安慰治疗组):10只,先制成HIE模型,然后给予PBS作为安慰治疗；SHAM组(假手术组):10只,仅按HIE模型制作过程施予假手术,不给予其他干预.干预后第7、14、21、28、35天用神经行为学实验Rotarod Test、Cylinder Test检测各组小鼠的行为学变化；干预后第1-5天及第2、6个月进行Morris水迷宫测试,比较结果的异同.结果 1)Rotarod Test:HIE+NSC组小鼠运动功能逐渐恢复,至第14天时与HIE+PBS组相比差异具有统计学意义(P＜0.05),第21天时已恢复至接近手术前的水平.2)Cylinder Test:HIE+ NSC组随着时间恢复,28 d后,HIE+ NSC组和HIE+ PBS组的缺血侧肢体的使用频率差异有统计学意义(P＜0.05).3) Morris水迷宫实验:3组小鼠的逃避潜伏期随训练天数增加而逐渐缩短,前5d的总体学习成绩的组间差异有统计学意义(P＜0.001).第6个月和第2个月相比较,HIE+ NSC组小鼠的逃避潜伏期缩短(P＜0.05),HIE+NSC组与HIE+ PBS组小鼠的逃避潜伏期在各个时间点的差异均有统计学意义(P＜0.05).结论 Rotarod Test主要适用检测急性期神经功能的障碍和恢复情况.Morris水迷宫可作为检测新生小鼠HIE经NSC治疗后期的行为学变化的指标.Cylinder Test是一种简单经济的神经行为学实验,适用于任何时期,也适合与其他行为学实验结合使用.%Objective To observe the changes in behaviors of hypoxic-ischemic encephalopathy (HIE) mice treated with neural stem cells (NSC),and to provide support for treating HIE with NSC,and to verify the feasibility of rotarod test,cylinder test and Morris water maze for measuring neurological deficit and recovery. Methods After HIE was induced, 30 7-day-old mice were randomly assigned to receive NSC (NSC group,n= 10) ,PBS (PBS group,n = 10) and sham operation (SHAM group,n=10). Rotarod Test and Cylinder Test were performed at 7,14,21,28 and 35 days after treatment and Morris water maze test was carried out at 1-5 days, and 2 and 6 months after treatment to measure the behaviors of mice. Results 1)Rotarod Test: motor func-tion recovered gradually and restored to near pre-operative levels on the 21th day in NSC group, and there was a significant difference between NSC group and PBS group on the 14th day (P< 0. 05). 2)Cylinder Test: motor function recovered with time in NSC group, and use frequency of ipsilateral forelimb was significantly different between NSC group and PBS group at 28 days after treatment (P<0. 05). 3) Morris water maze test-escape latency shortened gradually with increasing training days,and the differences were significant among the three groups during the first 5 days after treatment (P<0. 001). Compare with results at 2 months after treatment,escape latency obviously decreased at 6 months after treatment in NSC group(P<0. 05). Compared to PBS group,escape latency significantly shortened in NSC group at each time point(P<0. 05). Conclusion Rotarod Test is mainly applied to the detection of acute neurological disorder and recovery. Morris water maze test can be used to determine the changes in behaviors of HIE mice treated with NSC in the later period. Cylinder Test is a simple and economic experiment,which can be applied to neurobehavioral assessment in any period and may be used in conjunction with other behavioral test.