摘要
目的建立符合国际化的临床前实验标准的实验性自身免疫性重症肌无力(experimental autoimmune myasthenia gravis, EAMG)动物模型。方法参照文献报道的方法并改进后,从电鳐电器官提取乙酰胆碱受体(acetylcholine receptor,AchR)蛋白纯品,并采用SDS凝胶电泳蛋白定性鉴定及BCA法蛋白定量;用纯化的蛋白主动免疫C57BL/6小鼠,共免疫3次(分别于第1天、第30天、第60天),进行EAMG小鼠临床评分、体重、血清AchR抗体含量、新斯的明试验、肌电图等综合评价。结果 EAMG模型组与佐剂组比较,自第三周开始发病,平均临床评分显著上升(P<0.01);发病小鼠体重显著减轻(P<0.01);新斯的明试验阳性;血清AchR抗体含量明显增加(P<0.01);肌电图重复电刺激实验阳性。结论从黑斑双鳍电鳐的电器官提取、纯化AchR蛋白成功诱导C57BL/6 EAMG小鼠模型,为进一步研究重症肌无力创造了良好条件。
Objective To establish a mouse model of experimental autoimmune myasthenia gravis (EAMG) by immunization with purified AchR extracted from the Narcine maculata electric organ, which meets the requirements of international criteria of preclinical experiment. Methods Purified AchR protein was extracted from the electric organ of Narcine maculata and validated by SDS-polyacrylamide gel electrophoresis. C57BL/6 mice were immunized with the protein and complete Freund adjuvant (CFA) for the EAMG group or CFA alone for the control group three times at days 1, 30, and 60. After 70 days, the clinical assessments were determined according to body weight, serum anti-AchR antibody level, neostigmine text, and electromyography. Results Compared with control mice, the clinical scores of EAMG mice were increased significantly ( P < 0.01). Body weight was significantly decreased ( P < 0.01), neostigmine test was positive, and the percentage of decrement of the fifth wave of electromyography and levels of anti-AchR antibody were significantly higher ( P < 0.01). Conclusions The C57BL/6 EAMG mouse model is successfully established by immunization with purified AchR protein extracted from the Narcine maculata electric organ, which may provide a better research tool for studies of experimental autoimmune myasthenia gravis.
作者
李志彬
李毅
金宛霖
杨树梅
孟环宇
胡波
徐立群
罗朝辉
杨欢
LI Zhibin;LI Yi;JIN Wanlin;YANG Shumei;MENG Huanyu;HU Bo;XU Liqun;LUO Zhaohui;YANG Huan(Department of Neurology, Xiangya Hospital, Central South University, Changsha 410008, China)
出处
《中国实验动物学报》
CAS
CSCD
北大核心
2019年第2期143-147,共5页
Acta Laboratorium Animalis Scientia Sinica
基金
国家自然科学基金项目(81571173
81771364)~~
