miR-30a对实验性自身免疫性脑脊髓炎模型小鼠发病的作用  

Effect of miR-30a on pathogenesis in experimental autoimmune encephalomyelitis model mice

作　　者：刘守正[1] 王婷 张英 江红艳[1] 张智慧[1] 曲学彬 姚瑞芹 Liu Shouzheng Wang Ting Zhang Ying Jiang Hongyan Zhang Zhihui Qu Xuebin Yao Ruiqin(Orthopaedics Department, Shuyang People＇s Hospital, Shuyang 223600 Research Center of Neurobiology, Xuzhou Medical University, Xuzhou 221009, China)

机构地区：[1]沭阳县人民医院骨科,沭阳223600 [2]徐州医科大学神经生物学研究中心,徐州221009

出　　处：《解剖学杂志》2017年第1期7-10,共4页Chinese Journal of Anatomy

基　　金：国家自然科学基金(81271345;81302519);江苏省自然科学基金(BK20131132);江苏省2014高校年度"青蓝工程"中青年学术带头人资助项目

摘　　要：目的:探讨在体过表达miR-30a对实验性自身免疫性脑脊髓炎(EAE)模型小鼠脱髓鞘病变的作用。方法:构建含稳定表达miR-30a的shRNA慢病毒载体,以含有空载体的慢病毒为对照(LV-ctrl)。雌性C57BL/6小鼠随机分为正常对照组(normal)、对照慢病毒(LV-ctrl)组和miR-30a慢病毒(LV-miR-30a)组,2组慢病毒经尾静脉注射,注射7 d后用MOG35-55多肽免疫小鼠。Kono 5分法检测发病情况,快蓝(LFB)染色及透射电镜检测脊髓脱髓鞘程度及髓鞘结构的变化。结果:LV-ctrl组小鼠在MOG_(35-55)多肽免疫的第10天开始发病,到第21天,其Kono评分为2.57±0.79,LV-miR-30a组小鼠的Kono评分显著低于LV-ctrl组小鼠;miR-30a处理可显著提高小鼠体质量增长率,同时LFB评分也较LV-ctrl组显著增加;髓鞘超微结构显示LV-ctrl组小鼠脊髓髓鞘板层松解且稀疏,而LV-miR-30a组小鼠的髓鞘结构相对完好,髓鞘厚度显著大于LV-ctrl组。结论:过表达miR-30a可以明显减缓EAE的发病,减轻脊髓脱髓鞘程度。Objective： To investigate the effect of miR-30a overexpression on the pathogenesis and demyelination in experimental autoimmune encephalomyelitis （EAE） model mice. Methods： MiR-30a shRNA lentivirus was constructed and the lentivirus containing mutant gene sequence was selected as the control. Female C57BL/6 mice were randomly divided into 3 groups： normal control group （normal）, control lentivirus group （LV-ctrl） and miR-30a lentivirus group （LV-miR-30a） in which the mice were injected with controlled lentivirus or miR-30a lentivirus by caudal vein. Mice were immunized with MOG35-55 peptide 7 days after lentivirus injection. The disease condition was detected by Kono＇s 5-point. Luxol fast blue（LFB） staining and transmission electron microscopy were used to detect the degree of demyelination and the structure of myelin sheath. Results： LV-ctrl group mice began to fall ill on the tenth day after being immunized with MOGas_ss peptide. On the twenty-first day, the Kono＇s score in LV-ctrl group mice was 2.57 ± 0.79, and the score in the LV-miR-30a group was significantly lower than that in the LV-ctrl group. The growth rate of body weight was significantly enhanced in the LV-miR- 30a group compared with the LV-ctrl group. Meanwhile, miR-30a also increased the LFB score. The ultrastructure of myelin sheath showed that myelin sheath of the spinal cord in the LV-ctrl group was loose and sparse, while relatively intact in LV- miR-30a group; and the thickness of myelin sheath in LV-miR-30a group was much thicker than that in the LV-ctrl group. Conclusion： miR-30a over-expression in vivo can significantly inhibit the incidence of EAE, and relieved the demyelination.

关 键 词：脊髓 实验性自身免疫性脑脊髓炎 髓鞘 脱髓鞘 miR-30a 小鼠 

分 类 号：R-332[医药卫生] R744.51