B淋巴细胞刺激因子（BLyS）与实验性重症肌无力动物模型的相关性研究

发布时间：2018-04-20 15:33

本文选题：实验性自身免疫性重症肌无力 + 乙酸胆碱受体　；参考：《南京师范大学》2006年硕士论文

【摘要】：本文通过建立实验性自身免疫性重症肌无力动物(experimental autoimmune myasthenia gravis，EAMG)模型，利用抗BLyS抗体在EAMG模型建立前的预防和建立后的治疗试验，检测了抗BLyS抗体在EAMG中的预防和治疗作用，并初步探索了抗BLyS抗体在EAMG中的作用机理。结果如下： 1 实验性自身免疫性重症肌无力(EAMG)动物模型的建立与鉴定用烟碱型乙酰胆碱受体α1亚单位125～149多肽片段和鼠伤寒沙门氏裸菌免疫C57BL／6小鼠，观察小鼠的临床症状、抗体滴度、游泳疲劳时间及肌电图变化。结果表明：大部分模型组小鼠强化接种后出现轻微肌无力症状，级别评定为1分；模型组小鼠的抗体滴度与正常对照组小鼠差异显著，20只模型组小鼠中有12只的抗体滴度为阳性；游泳试验中有18只小鼠呈阳性；肌电图检查中所有小鼠均为阳性，最大衰减幅度达82％。提示：成功建立了EAMG模型，为进一步研究EAMG的发病机理打下了基础。 2 抗BLyS和BSA多克隆抗体的制备和纯化选取2-3kg的雄性日本大耳白兔，分别采用乆淋巴结免疫法和皮下免疫法免疫兔子，制备抗血清；间接ELISA法检测抗血清的效价；并应用分级饱和硫酸铵盐析法进行纯化。结果表明：获得抗BLyS多克隆抗体，间接ELISA法检测抗血清的效价为50万倍；获得抗BSA多克隆抗体，间接ELISA法检测抗血清的效价为5万倍。提示：分别成功制备了抗BLyS和抗BSA多克隆抗体，为进一步研究抗BLyS和重症肌无力动物模型之间的相关性奠定了基础。 3 抗BLyS抗体对EAMG的预防和治疗作用研究建立实验性自身免疫性重症肌无力动物预防组、预防对照组、治疗组和治疗对照组，比较四组小鼠的临床症状、抗体滴度、游泳疲劳时间及肌电图变化。结果表明：预防组小鼠的临床症状评为0分，抗体滴度80％为阴性，游泳疲劳时间为阴性，肌电图衰减均为阴性，，与正常组小鼠之间的差异不显著；治疗组临床症状评为1分，抗体滴度60％为阳性，40％为可疑阳性，游泳疲劳时间为阳性，肌电图衰减均为阳性，与正常组小鼠之间的差异显著；预防对照组和治疗
[Abstract]:In this study, we established an experimental autoimmune myasthenia gravis animal (experimental autoimmune).
Myasthenia gravis, EAMG) model, using the prevention and establishment of anti BLyS antibody before the establishment of the EAMG model, the preventive and therapeutic effects of anti BLyS antibody in EAMG are detected, and the mechanism of anti BLyS antibody in EAMG is preliminarily explored. The results are as follows:
1 Establishment and identification of experimental autoimmune myasthenia gravis (EAMG) animal model
C57BL / 6 mice were immunized with nicotinic acetylcholine receptor alpha 1 subunit 125~149 polypeptide and mice with Salmonella typhimurium. The clinical symptoms, antibody titers, swimming fatigue time and electromyography were observed in mice. The results showed that most of the model mice had mild myasthenia symptoms after intensive inoculation, and the grade evaluation was 1. The antibody titer of the mice in the type group was significantly different from that of the normal control group. In the 20 model group, 12 of the mice were positive, and 18 mice were positive in the swimming test. All the mice were positive in the electromyography examination. The maximum attenuation range was 82%.: the EAMG model was built successfully to further study the pathogenesis of EAMG. The mechanism lays the foundation.
Preparation and purification of 2 anti BLyS and BSA polyclonal antibodies
The antiserum was prepared by immunization with lymph node immunization and subcutaneous immunization, and the titer of antiserum was detected by indirect ELISA method, and the anti BLyS polyclonal antibody was obtained. The results showed that the anti BLyS polyclonal antibody was obtained and the titer of the antiserum was detected by indirect ELISA method. 500 thousand times, the anti BSA polyclonal antibody was obtained, and the titer of the indirect ELISA assay was 50 thousand times. It was suggested that the anti BLyS and anti BSA polyclonal antibodies were successfully prepared, which laid the foundation for further study on the correlation between anti BLyS and myasthenia gravis model.
3 the effect of anti BLyS antibody on the prevention and treatment of EAMG
The experimental autoimmune myasthenia gravis group was established to prevent the control group, the treatment group and the treatment control group. The clinical symptoms, antibody titers, swimming fatigue time and electromyography of the four groups were compared. The results showed that the clinical symptoms of the prevention group were 0, the antibody titer 80% was negative, and the swimming fatigue time was negative. The EMG attenuation was negative, and the difference between the normal group and the normal group was not significant. The clinical symptoms of the treatment group were 1 points, the antibody titer 60% was positive, the 40% was suspicious positive, the swimming fatigue time was positive, the EMG attenuation was all positive, and the difference between the normal group and the normal group was significant; the prevention control group and the treatment group were treated and treated.

【学位授予单位】：南京师范大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392;R-332

【参考文献】