实验性自身免疫性脑脊髓炎对大脑神经干细胞增殖的影响  

Effect on neural stem cell proliferation in experimental autoimmune encephalomyelitis

作　　者：郭俊[1] 李宏增[1] 林宏[1] 苗建亭[1] 宿长军[1] 李柱一[1] 

机构地区：[1]第四军医大学唐都医院神经内科,710038

出　　处：《中国神经免疫学和神经病学杂志》2010年第4期238-241,246,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：国家自然科学基金资助项目(30570641)

摘　　要：目的探讨实验性自身免疫性脑脊髓炎(EAE)埘大脑神经干细胞(NSC)增殖能力的影响。方法将C57BL/6小鼠随机分为模型组、佐剂组和对照组,采用少突胶质细胞糖蛋白(MOG)抗原联合完全福氏佐剂(CFA)免疫模型组小鼠,佐剂组小鼠仅接受CFA免疫注射,对照组小鼠仅接受生理盐水注射。观察各组小鼠EAE症状评分,采用HE染色以及劳克坚牢蓝(LFB)髓鞘染色观察其脑和脊髓病理学改变,采用免疫荧光染色检测小鼠海马齿状回(DG)和侧脑室下区(SVZ)BrdU阳性细胞数量,以评价NSC增殖情况。结果模型组有14只(14/15)小鼠被成功诱导EAE,其症状达到高峰后短暂缓解,随后进入慢性持续期;而佐剂组和对照组小鼠均未出现任何EAE症状。模型组中枢神经系统存在大量炎性反应细胞浸润,在白质区可见多处LFB未着色的髓鞘脱失区;而佐剂组和对照组无明显病理学改变。佐剂组与对照组小鼠DG区和SVZ区BrdU阳性细胞数比较差异均无统计学意义。与佐剂组和对照组比较,模型组小鼠大脑SVZ区和DG颗粒下区BrdU阳性细胞均显著降低(均P<0.05)。结论慢性持续型EAE可导致小鼠大脑NSC增殖能力下降。Objective To explore the change of neural stem cell （NSC） proliferation in experimental autoimmune encephalomyelitis （EAE）. Methods C57BL/6 mice were randomly divided into model group, CFA group and control group. EAE was induced by subcutaneous immunization with 200 vg of MOG35-55 peptide in complete Freund＇ s adjuvant （CFA） containing 4 mg/mL myeobacterium tuberculosis H37Ra at four sites in the back or the flank of each mouse; A fixed amount （200 ng） of pertussis toxin was intraperitoneally （i. p. ） administered at 0 and 2 days post-immunization （dpi）. Meanwhile, CFA-immunized mice underwent a subcutaneous immunization with CFA and two i. p. injections of pertussis toxin; Control mice only received 0.9% saline （n=15 per group）. EAE mice were determined by assessment of clinical signs of paralysis. Pathological changes in the CNS were evaluated by H＆E and luxol fast blue （LFB） staining. At 21 dpi, mice from the three groups received two i.p. injections of 5-bromo-2＇ -deoxyuridine （BrdU） 24 h before they were killed. Coronal brain sections were collected and stained with anti-BrdU antibody. The NSC proliferation was assessed by counting the number of BrdU-positive cells in the dentate gyrus （DG） of the hippocampal formation and in the subventricular zone （SVZ） of the lateval cerebroventricle, respectively. Results EAE was successfully developed in 14 of 15 MOG-immunized mice, while no signs of paralysis occurred in both CFA-immunized and control mice. In EAE mice, the peak of disease was followed by a brief remission and then a stable chronic phase. H＆E staining showed that inflammatory infiltration was present in the CNS of EAE mice; areas of myelin loss were observed in white matters of the CNS by LFB staining. In contrast, no pathological changes present in CFA- immunized and control mice. For detection of NSC proliferation, 6 mice were randomly chosen from each group, respectively, and were injected with BrdU at 21 dpi. Compared with CFA-immunized and contr

关 键 词：脑脊髓炎 实验性自身免疫性 神经干细胞 增殖 

分 类 号：R744.51[医药卫生—神经病学与精神病学]