中图分类号R743.33文献标识码A文章编号1007-5496(2000)05-0678-02
Change of histo- morphology on the rats' cerebral infarction after rehabilitative training
XU Li
(XiJing Hospital, Fourth Military Medical University,Xi'an 710032,China)
LI Ling
(XiJing Hospital, Fourth Military Medical University,Xi'an 710032,China)
YAN Pei-song,et al.
(XiJing Hospital, Fourth Military Medical University,Xi'an 710032,China)
Abstract Objective:To study the change of histo- morphology on the rats' cerebral infarction after rehabilitative training. Methods:65 SD rats were given cerebral infarction induced by photochemical injure and were divided into 3 groups at random: rehabilitation training group,braking group and free group after 24 hours.Rehabilitation group which were given balancing,grasping,rotating and walking exercises everyday, braking group which were fixed in mesh cages and free group which were feeded in normal cages, three groups were measured volume of infarction tissue and obsevred change of morphology microscope at 24 h,1 week,2 week,3 week and 4week. Results: Every groups were the largest volume of brain infarct at 24 h and obviously reduced at 1 week,especially in rehabilitation group(P<0.05),then it have gradually decreased and not have difference at 2~ 4 week. Microscope showed obviously necrosis inside brain infarct focal at 24 h . Rehabilitation groups showed that glia cells had hyperplasia in edge of brain infarct and a lot of blood vessel sprout had grown into brain infarction at 1 week.then it formed scar at 2 week and the edge of brain infarction focal had more hyperplasia of fibroblast obviously than the free group and braking group at 3 week.The granultion tissue and bracket of blood vessel formed in the brain infarction focal of 4 week ,blood vessel had hyperplasia and dilatation in the free group and braking group.Conclusions:Rehabilitative training can reduce volume of infarction early and promot blood circulation and improved ischemia and regeneration and repair of brain tissue.
Key words rehabilitation;rats;cerebral infarct;histo-morphology
脑血管病严重危害人类的健康,近年来,对于脑血管病的康复研究越来越受到人们的重视,为了提高生活质量,减少残疾,重返社会,康复功能锻炼已作为脑血管病恢复中不可缺少的手段。康复治疗对缺血性脑梗塞患者在改善感觉、运动、行为能力方面已获得明显的疗效[1,2];为了观察康复后脑梗塞组织结构的改变;在实验中,我们采用CPD4作光敏剂,氦氖(He-Ne)激光作光源,制做光化学诱导的脑梗塞模型,进行康复功能训练,在24h、1周、2周、3周、4周给于测量脑梗塞体积及光镜观察病理学方面的改变,以了解其恢复状况。
1材料和方法
1.1实验材料
1.1.1材料CPD4粉剂由杭州民生药厂提供,批号930502于0~4℃避光保存,使用前用生理盐水稀释至所需浓度。
1.1.2动物雄性SD大鼠,由第四军医大学实验动物研究中心提供,体重(240±50)g,鼠龄12周左右,二级动物。
1.1.3光源采用西安激光仪器厂生产的795B型氦氖激光双管治疗机,输出波长6328Å(可见红光),采用原光束双管照射,输出功率8mW。
1.1.4康复评估及训练器材采用自制滚筒式网状训练器;其中长100cm、直径60cm的圆形网状仪器;中间被分为4个格;可同时训练4只鼠;底座有一固定架;一端有一手摇柄;可手摇按每分钟5转进行转动训练;该器材可训练大鼠的抓握、旋转、行走等运动。平衡木训练采用了170cm长、2cm宽的方木棒,平放在距地面7cm处,作为一个平衡木让鼠在其上行走,主要评估及训练平衡功能。转棒训练取长150cm,直径4.5cm的木棒1根,其中点固定在3转/分的转动器上,分别向左右交替转动,可评估及训练动态的平衡功能。网屏训练:网屏为50cm×40cm网带,网眼为1cm×1cm,网板的左右和上方都用25cm高的木板框边,网屏距地面高度为80cm,下方铺以12cm厚的海绵,先将网屏水平放置,将老鼠放在其上,然后缓缓地将其一端抬高,在2s内将此屏风变成垂直位,保持5s,观察大白鼠是否会从网屏上掉下来或用前爪握住网屏。从而客观地评价前爪抓握能力及肌力情况。
1.2实验方法
1.2.1动物分组65只大鼠造模后随机分3组:康复训练组,每天给予平衡、抓握力、旋转、行走等训练。自由活动组,置于普通笼内饲养,可自由活动。制动组,置于长40cm、直径6cm的网状笼内固定,在头端有一容器给予食物和水,四肢和身体处于固定状态。3组分别于造模后24h、1周、2周、3周、4周各取5只,给予TTC染色、HE染色测量梗塞面积及光镜检察。
1.2.2模型制做方法大鼠1%戊巴比妥钠(0.3ml/100g)腹腔注射麻醉,健侧股静脉注射CPD4后10min内,将鼠俯卧位于WDT-V型大鼠头颅定位仪(西安西北光学仪器厂生产),沿头颅正中线偏右切开头皮暴露颅骨前沿部位,在前囟门后1.8mm,正中线旁开约2mm,部位为上下肢皮层感觉运动区,用黑色避光纸(中央有一8mm×5mm的小孔)遮避周围组织,He-Ne激光照射20min,术后缝合头皮,避光24h。
1.2.3观察、取材及处理实验后24h、1周、2周、3周、4周后各组进行断头取脑,去除嗅部、小脑、低位脑干,在视交叉前后等分间隔2mm连续作6个脑冠状切片,将其放入提前预热的37°C2%TTC生理盐水溶液中30min,染色后正常组织为红色,脑梗塞组织为白色,将脑片平面置于刻度纸上,用数码相机照相后输入计算机,采用Photoship软件计算每片梗塞面积;然后按公式V=Sd/T2(S为每一脑片梗塞灶面积,d为脑片厚度,T为线形放大倍数)计算每一脑梗塞体积(图2);最后取材放入10%福尔马林液固定,石蜡包埋,用切片机5μm切片,HE染色,光镜下观察组织学变化。
图2脑梗塞体积模型示意图
1.3统计学处理采用Origin统计软件进行统计分析,按±s作统计图,按t检验进行两两比较。
2结果
2.1脑梗塞体积24h脑梗塞体积最大,1周后时各组明显减少,尤以康复组为著(P<0.05),2~4周体积逐渐减小,而且各组间无明显差异。(详见图1)。
图1各组不同时间脑梗塞体积的变化
2.2脑组织病理学改变HE染色,光镜下24h可见脑梗塞灶区神经元细胞有肿胀、细胞核固缩、碎裂,有的溶解、消失;胞浆变红,严重区胶质细胞消失;脑梗塞灶周边区HE染色变浅,与坏死区界线清楚。1周时梗塞灶区有脑细胞软化,胶质细胞溶解及大量的泡漠细胞,仍有少量细胞核固缩及小的毛细血管扩张,增生;灶边缘有胶质细胞增生,尤以康复组增生为著,并且在梗塞灶周边区有散在的小血管芽向坏死灶区生长,制动组、自由组灶边缘胶质细胞增生少,未见小的血管芽。2周时梗塞灶区有急、慢性炎细胞浸润及纤维母细胞增生;梗塞灶边缘有胶质细胞增生,毛细血管扩张,康复组梗塞灶边缘形成胶质瘢痕,而制动组、自由组灶边缘胶质细胞增生少,无胶质瘢痕形成;3周梗塞灶内有大量中性白血病浸润,中央有液化性坏死,梗塞灶周边境界清楚、局限,康复组梗塞灶边缘仍有血管、纤维母细胞增生;制
