摘要
目的观察不同治疗方法下EAE小鼠神经组织及少突胶质前体细胞的不同病理阶段下的形态及变化。方法将100只雄性小鼠随机分成对照组、造模组、β-干扰素组、胚胎鼠OPCs移植组,并采用Brdu腹腔注射。(1)对照组小鼠于0、7 d皮下注射生理盐水+CFA免疫乳剂0.2 ml,于0、48 h尾部静脉注射百日咳毒素。(2)造模组、β-干扰素组、胚胎鼠OPCs移植组小鼠均于0,7 d皮下注射MOG+CFA免疫乳剂0.2 ml,于0、48 h尾部静脉注射百日咳毒素。β-干扰素组小鼠第8天开始皮下注射INFβ-1a 1μg,连续14 d。胚胎鼠OPCs移植组小鼠并于第1天、第3天和第7天分别于侧脑室注射移植OPCs。分别于模型诱导后第14天、21、28、35天处死EAE小鼠(每次每组各处死5只),取其脑组织及脊髓连续薄层切片,分别进行HE染色、Luxol Fast Blue髓鞘染色等,观察其神经病理和免疫组化改变,并观察NCAM(+)和Brd U(+)双标记物阳性免疫荧光染色结果,同时进行电镜观察的标本取材,进行电镜观察。结果除对照组外其他三组神经功能评分在造模后均呈先上升再下降趋势,在第28天时神经功能评分最高,同时胚胎鼠OPCs移植组在第21、28、35天时神经功能评分均低于β-干扰素组和对照组(P<0.05)。对照组小鼠在各时间段侧脑室外侧壁和脊髓腹侧白质的NCAM(+)、Brd U(+)及GFAP(+)、Brd U(+)无显著变化,但其他三组NCAM(+)、Brd U(+)及GFAP(+)、Brd U(+)的阳性表达均在第28天时达到最高(P<0.05),随后下降但仍高于对照组(P<0.05)。胚胎鼠OPCs移植组在第21、28、35天时其NCAM(+)、Brd U(+)均高于β-干扰素组和对照组(P<0.05)。结论少突胶质前体细胞在髓鞘重建过程中起重要作用,在不同修复阶段具有不同形态,且胚胎鼠OPCs移植法可显著增加体内少突胶质前体细胞的量,值得推广使用。
Objective To observe oligodendrocytes morphology in EAE mice in different pathological stages. Methods 100 male mice were randomly divided into control group,building modules,beta interferon group,the embryonic rat OPCs transplantation group,and USES the Brdu intraperitoneal injection. The control mice were injected with normal saline ＋ CFA immunoemulsion 0. 2 ml in 0,7 d,and intravenous injection of pertussis toxin at the end of 0,48 h. Mice in the experimental group were injected with 0. 2 ml of MOG ＋ CFA immunoemulsion at 0,7d,and injection of pertussis toxin at the tail vein of 0,48 h. The eighth day the mice in the beta interferon group were injected with the subcutaneous injection of INF beta-1a 1 μg for 14 consecutive days. The OPCs were transplanted from the lateral ventricle on day 1,3 and 7 respectively in model induction day 14,21,28,35 days after death EAE mice（ each executed only 5 in each group）,take its continuous thin layer of the brain and spinal cord slice,for HE staining,Luxol Fast Blue myelin,dyeing,etc. Observed the pathological and immunohistochemical changes,its nerve and observe NCAM（ ＋） and Brdu positive immunofluorescence staining（ ＋） double markers as a result,the electron microscope samples of materials at the same time,the electron microscope. Results Except for the control group,nerve function score in the other three groups after building raised by first and then decline,at the same time in the day 28 nerve function scored the highest,embryonic rat OPCs transplantation group in 21,28,35 days their nerve function scored lower than beta interferon group and the control group（ P〈0. 05）. NCAM（ ＋）,Brd U（ ＋） and GFAP（ ＋）,Brd U（ ＋） of the lateral ventricle wall and spinal cord ventral white matter in control group in each period had no significant change,but NCAM（ ＋）,Brd U（ ＋） and GFAP（ ＋）,Brd U positive expression of（ ＋） in the other three groups reached the highest in28 days（ P〈0. 05）,and then drop but still higher than the control group（ P〈0. 05）. At the same time NCAM（ ＋）,Brd U（ ＋） of embryonic rat OPCs transplantation group in 21,28,35 days were higher than the beta interferon group and the control group（ P〈0. 05）. Conclusion Oligodendrocyte precursor cell plays an important role in the process of the myelin sheath reconstruction,has different forms in different restoration stage,and embryonic rat OPCs transplants can significantly increase the less amount of glial progenitor cells in the body. Therefore,it is worth clinical promotion.
出处
《临床和实验医学杂志》
2017年第19期1884-1889,共6页
Journal of Clinical and Experimental Medicine
基金
湖北省自然科学基金项目(2015CFB476)
关键词
小鼠
多发性硬化
少突胶质前体细胞
髓鞘重建
Mice
Multiple sclerosis
Oligodendrocyte precursor cell
Myelin sheath reconstruction
