LINGO-1基因沉默对自身免疫性脑脊髓炎小鼠运动功能的影响  

Effect of LINGO-1 gene silencing on movement function of EAE mice

作　　者：郭守刚[1] 张杰[2] 汪春娟[1] 杜怡峰[1] 

机构地区：[1]山东大学附属省立医院神经内科,济南250021 [2]山东省邹平县人民医院神经内科,济南250021

出　　处：《中华医学杂志》2015年第39期3223-3228,共6页National Medical Journal of China

基　　金：国家自然科学基金（81171171,30970991,81100802）;山东省中青年科学家奖励基金（博士基金）（2014BSB14119）

摘　　要：目的 探讨沉默LINGO-1基因表达对自身免疫性脑脊髓炎（EAE）小鼠运动功能的影响.方法 通过MOG35-55诱导雌性C57/BL6小鼠建立EAE小鼠模型.将EAE小鼠（105只）完全随机分为5组：A组（21只）：侧脑室注入5μl滴度为5×109 Tu/ml编码LINGO-1shRNA的慢病毒载体（LV/LINGO-1-shRNA）;B组（21只）：侧脑室注入5μl滴度为5×10s Tu/ml LV/LINGO-1-shRNA;C组（21只）：侧脑室注入5μl滴度为5×107 Tu/ml LV/LINGO-1-shRNA;D组（21只）：侧脑室注入5μl无义序列的LVCON053;E组（21只）：未治疗组.动态观察干预后不同时间点小鼠神经功能评分情况及LINGO-1蛋白表达水平,并行髓鞘快蓝染色了解各组髓鞘脱失情况.结果 侧脑室注射LV/LINGO-1-shRNA组EAE小鼠脑组织内LINGO-1蛋白表达从干预后第7天起可见明显下调,其中以B组和C组下调最明显（1.99±0.13,2.08±0.10,P＜0.05,P＜0.01）.从第7天开始,相比较于D组和E组,A组、B组和C组运动功能评分有不同程度地下降[（3.11 ±0.13）分,（2.42 ±0.13）分,（2.96 ±0.10）分比（3.56 ±0.15）分,（3.87 ±0.12）分,P＜0.01,P＜0.01,P＜0.05].30 d时A组和B组髓鞘密度明显高于未治疗组（0.72±0.09,0.83 ±0.11比0.56±0.10,P＜0.05,P＜0.01）.结论 侧脑室注射LV/LINGO-1 shRNA是一种沉默EAE小鼠脑内LINGO-1表达的有效方式.LINGO-1下调可改善EAE小鼠运动功能及促髓鞘修复,但效果并不随LV/LINGO-1-shRNA剂量增加而增强.Objective To explore the effect of LINGO-1 silencing on movement function of experimental autoimmune encephalomyelitis （EAE） mice.Methods EAE was established by induction of MOG35-55 in female C57/BL6 mice.Then female EAE mice （n =105） were completely randomly divided into 5 groups：group A （n =21） ：5 μl 5 × 109 Tu/ml lentiviral vectors encoding LINGO-1shRNA （LV/ LINGO-1-shRNA） by intracerebroventricular （ICV） injection, group B （n =21） ：5 μl 5 × 108Tu/ml LV/LINGO-1-shRNA by ICV injection, group C （n =21） ： 5 μl 5 × 107 Tu/ml LV/LINGO-1-shRNA by ICV injection,group D （n =21）：5 μl LVCON053 by ICV injection and group E （n =21）：untreated.The movement function was scored and the expression of LINGO-1 protein was detected by Western blot on day 1,3,7,14,21,30 after ICV among different groups.Luxol fast blue staining was performed to know about conditions of myelin sheath on day 30.Results The expression of LINGO-1 in EAE mouse was obviously downregulated ever since day 7 after LV/LINGO-1-shRNA implantation.Group B and C achieved the most reduction of LINGO-1 expression （1.99 ± 0.13,2.08 ± 0.10, P 〈 0.05, P 〈 0.01）.Simultaneously, the movement functional score of group A, B and C was lowered at different levels from day 7 （3.11 ± 0.13, 2.42 ±0.13,2.96 ±0.10 vs 3.56 ±0.15,3.87 ±0.12,P 〈 0.01,P 〈0.01, P 〈0.05） ,with the most marked decrease in group B.The densities of myelin sheaths in group A and B were higher than untreated group on day 30 （0.72 ± 0.09,0.83 ± 0.11 vs 0.56 ± 0.10, P 〈 0.05, P 〈 0.01）.Conclusion LV/ LINGO-1shRNA by ICV injection is an effective method to silence LINGO-1 expression.LINGO-1 silencing could ameliorate motor function and promote formation of myelin sheaths.But the effects do not enhance with the increase of LV/LINGO-1-shRNA dose.

关 键 词：实验性自身免疫性脑脊髓炎 LINGO-1 基因沉默 髓鞘 

分 类 号：R744.5[医药卫生—神经病学与精神病学]