骨髓间充质干细胞移植后脑梗死大鼠脑电图的变化

Alterations of electroencephalogram in rats with cerebral infarction after the transplantation of bone marrow mesenchymal stem cells

目的:对比观察骨髓间充质干细胞移植前后脑梗死大鼠脑电图的变化。方法:实验于2002-09/12在解放军第三军医大学中心实验室及西南医院神经内科肌电图室完成。①实验分组:选取清洁级健康成年Wistar大鼠15只,随机数字表法分为干细胞移植组、模型对照组、假手术组,5只/组。②实验方法:另取2只健康幼年Wistar大鼠用于骨髓间充质干细胞的提取,联合采用密度梯度离心及贴壁法分离培养大鼠骨髓间充质干细胞,选取生长良好的1～3代细胞用于移植实验。干细胞移植组、模型对照组大鼠建立大脑中动脉栓塞模型。假手术组仅分离颈总动脉、颈外动脉和颈内动脉,不予结扎和放置线栓。造模后1周,干细胞移植组、假手术组大鼠行细胞移植,在立体定向仪定位下于脑梗死区(壳核)直接注射骨髓间充质干细胞悬液5μL,细胞浓度1×104μL-1,移植坐标为前囟前1.0mm,右旁开3.0mm,硬膜下5.0mm。模型对照组大鼠于相同部位注射等量不含细胞的磷酸盐缓冲液。③实验评估:采用脑电图机分别于造模前、造模后1周(移植前)、细胞移植后4周对各组大鼠进行脑电图检测。结果:15只大鼠均进入结果分析。①造模前基本节律为8～11Hz、15～30μV的α波,间或少量θ波,双侧对称。②造模后1周,假手术组异常率为0;模型对照组20%(1/5)轻度异常,80%(4/5)中度异常;干细胞移植组20%(1/5)轻度异常,60%(3/5)中度异常,20%(1/5)重度异常。③细胞移植后4周,假手术组脑电图恢复正常;模型对照组随术后时间的延长慢波有所减少,但仍可见到δ波、棘波、棘慢波的发放,至细胞移植后4周60%(3/5)轻度异常,40%(2/5)中度异常;干细胞移植组术后局限性慢波逐渐减少,基本节律全部恢复为α波,不对称的情况明显好转,至细胞移植后4周60%(3/5)轻度异常,以病灶侧局限性θ波较多为主,另外40%(2/5)基本正常。结论:动物实验显示骨髓间充质干细胞移植对脑梗死大鼠的脑电图背景节律有改善作用,一定程度上促进了神经系统功能的恢复。

AIM： To observe the changes of electroencephalogram （EEG） before and after the transplantation of bone marrow mesenchymal stem cells （MSCs） into the rats with cerebral infarction. METHODS： The experiment was conducted in the Central Laboratory and Laboratory of Electromyologram of South-western Hospital, Third Military Medical Hospital of Chinese PLA from September to December 2002. ①Fifteen healthy adult Wistar rats of clean grade were selected and randomly divided into MSCs transplantation group, model control group and sham operation group with 5 animals in each group. ②MSCs were isolated from another 2 healthy young Wistar rats by density gradient centrifugation methods, and cultured by attachment method. The first to third generation cells that grew well were selected. Rat models of middle cerebral artery occlusion （MCAO） were established in the former two groups, but in the sham operation group, the common carotid artery, external and internal carotid artery were isolated but not ligated. One week after modeling, 5 μL MSCs of 1×10^4 μL-1 were injected into the infarcted region （putamen） of the transplantation group and sham operation group, 1.0 mm anterior from bregma, right 3.0 ram, and 5.0 mm below dura mater. Meanwhile, the same volume cell-free phosphate buffer was injected into the rats of the control group. ③The examination of EEG was performed before, 1 week （before transplantation） after MCAO and 4 weeks after transplantation. RESULTS： All 15 rats were involved in the result analysis. ①Before modeling, the α wave of basic rhythm was 8-11 Hz and 15-30 μV, sometimes with few θ waves, and the two sides were symmetric. ②One week after modeling, the abnormal rate of the sham operation group was 0; 20% （1/5） of the model group were mild abnormity, and 80% （4/5） were moderate abnormity; 20% （1/5） of the transplantation group were mild abnormity, 60% （3/5） were moderate abnormity, and 20% （1/5） were severe abnormity.③Four weeks after MSCs transplantation, the EEG of the sham operation group was recovered; in the model group, the slow waves rate was decreased along with time, but still with wave, V wave and V slow wave, and 60% （3/5） were mild abnormity and 40% （2/5） were moderate abnormity 4 weeks after transplantation; in the transplantation group, the local slow waves were gradually decreased, all basic rhythm was wave, and unsymmetry was significantly improved. Four weeks after cell transplantation, 60% （3/5） were mild abnormity with local θ waves at focus side and 40% （2/5） were normal. CONCLUSION： The animal experiment shows that MSCs transplantation might improve the EEG of infracted rats and promote the recovery of neural function to some extent.