骨髓间充质干细胞移植急性一氧化碳中毒迟发性脑病大鼠星形胶质细胞及髓鞘的变化

背景：国内有关一氧化碳中毒迟发性脑病治疗的报道,多采用药物治疗和高压氧治疗,但存在治疗疗程长,见效慢,花费较高等问题。 目的：首次观察骨髓间充质干细胞移植治疗一氧化碳中毒迟发性脑病前后星形胶质细胞及神经髓鞘的变化。 方法：以随机抽签方法将SD大鼠分为3组：对照组、假手术组及干细胞移植组。采用腹腔注入一氧化碳方法制作一氧化碳中毒迟发性脑病模型,干细胞移植组在注射后第0,3,6,12,24,72小时及1周时将同种异体骨髓间充质干细胞经左侧颈动脉植入到大鼠脑内;假手术组扎闭颈外动脉,用PBS代替细胞悬液;对照组不进行细胞移植。移植后1,2,3,4周采用免疫组织化学的方法检测胶质纤维酸性蛋白和固绿-FCF染色法观察髓鞘着色情况。 结果与结论：造模后6,12,24 h干细胞移植组大鼠脑内胶质纤维酸性蛋白表达明显高于对照组及假手术组(P < 0.05),干细胞移植组移植后1~4周大鼠脑内胶质纤维酸性蛋白表达差异无显著性意义(P > 0.05)。造模后6,12,24 h干细胞移植组大鼠脑内髓鞘平均吸光度明显高于对照组及假手术组(P< 0.05),且细胞移植后第2周明显高于第1,3,4周(P < 0.05)。 提示经左侧颈动脉在一氧化碳中毒迟发性脑病大鼠体内移植骨髓间充质干细胞可以增加星形胶质细胞反应性,促进髓鞘再生,移植的最佳时机为发病后6~24 h,细胞移植后的积极作用在一两周最明显。

Changes in astrocytes and myelin sheath in rat models of delayed encephalopathy due to acute carbon monoxide poisoning following bone marrow mesenchymal stem cells transplantation

BACKGROUND:The therapy of delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) in China is drug or hyperbaric oxygen. They all cost higher and have long low effect. OBJECTIVE:To first observe the changes of astrocytes and nerve myelin before and after bone marrow mesenchymal stem cells (BMSCs) transplantation to treat the DEACMP. METHODS:Sprague-Dawley rats were divided randomly into three groups: control group, sham operated group, and stem cell transplantation group. All rats were injected carbon monoxide into abdominal cavity to produce DEACMP models. The transplantation group was injected with allograft bone marrow stromal cells through the left carotid artery at 0, 3, 6, 12, 24, 72 hours, 1 week after model establishment. External carotid artery was occluded in the sham operated group. The sham operated group was injected the same amount of phosphate buffered saline, and the control group was not injected anything. We detected the changes of glial fibrillary acidic protein (GFAP) by immunohistochemistry and observed myelin through Fast Green-FCF staining at 1, 2, 3 and 4 weeks posttransplantation.  RESULTS AND CONCLUSION:The expression of GFAP in the rat brain in stem cell transplantation group was significantly higher than that of control group and the sham operated group at 6, 12 and 24 hours (P < 0.05). No significant difference in GFAP expression of each group was detected at 1-4 weeks (P > 0.05). The average absorbance of myelin in stem cell transplantation group was significantly higher at 6, 12 and 24 hours than control and sham operated groups (P < 0.05); the average absorbance of myelin of stem cell transplantation group at 2 weeks were significantly higher than that at 1, 3, 4 weeks (P < 0.05). These indicated that transplanted BMSCs into the DEACMP rats models through the left carotid artery could increase the responsivity of GFAP and enhance remyelination. The best time for transplantation was between the 6 hours and the 24 hours and the positive effects were significantly observed between 1 week and 2 week after transplantation.