本文選題：實(shí)驗(yàn)性自身免疫性重癥肌無力 + 乙酸膽堿受體�。� 參考：《南京師范大學(xué)》2006年碩士論文

【摘要】：本文通過建立實(shí)驗(yàn)性自身免疫性重癥肌無力動(dòng)物(experimental autoimmune myasthenia gravis，EAMG)模型，利用抗BLyS抗體在EAMG模型建立前的預(yù)防和建立后的治療試驗(yàn)，檢測了抗BLyS抗體在EAMG中的預(yù)防和治療作用，并初步探索了抗BLyS抗體在EAMG中的作用機(jī)理。結(jié)果如下： 1 實(shí)驗(yàn)性自身免疫性重癥肌無力(EAMG)動(dòng)物模型的建立與鑒定 用煙堿型乙酰膽堿受體α1亞單位125～149多肽片段和鼠傷寒沙門氏裸菌免疫C57BL／6小鼠，觀察小鼠的臨床癥狀、抗體滴度、游泳疲勞時(shí)間及肌電圖變化。結(jié)果表明：大部分模型組小鼠強(qiáng)化接種后出現(xiàn)輕微肌無力癥狀，級(jí)別評(píng)定為1分；模型組小鼠的抗體滴度與正常對(duì)照組小鼠差異顯著，20只模型組小鼠中有12只的抗體滴度為陽性；游泳試驗(yàn)中有18只小鼠呈陽性；肌電圖檢查中所有小鼠均為陽性，最大衰減幅度達(dá)82％。提示：成功建立了EAMG模型，為進(jìn)一步研究EAMG的發(fā)病機(jī)理打下了基礎(chǔ)。 2 抗BLyS和BSA多克隆抗體的制備和純化 選取2-3kg的雄性日本大耳白兔，分別采用乆淋巴結(jié)免疫法和皮下免疫法免疫兔子，制備抗血清；間接ELISA法檢測抗血清的效價(jià)；并應(yīng)用分級(jí)飽和硫酸銨鹽析法進(jìn)行純化。結(jié)果表明：獲得抗BLyS多克隆抗體，間接ELISA法檢測抗血清的效價(jià)為50萬倍；獲得抗BSA多克隆抗體，間接ELISA法檢測抗血清的效價(jià)為5萬倍。提示：分別成功制備了抗BLyS和抗BSA多克隆抗體，為進(jìn)一步研究抗BLyS和重癥肌無力動(dòng)物模型之間的相關(guān)性奠定了基礎(chǔ)。 3 抗BLyS抗體對(duì)EAMG的預(yù)防和治療作用研究 建立實(shí)驗(yàn)性自身免疫性重癥肌無力動(dòng)物預(yù)防組、預(yù)防對(duì)照組、治療組和治療對(duì)照組，比較四組小鼠的臨床癥狀、抗體滴度、游泳疲勞時(shí)間及肌電圖變化。結(jié)果表明：預(yù)防組小鼠的臨床癥狀評(píng)為0分，抗體滴度80％為陰性，游泳疲勞時(shí)間為陰性，肌電圖衰減均為陰性，，與正常組小鼠之間的差異不顯著；治療組臨床癥狀評(píng)為1分，抗體滴度60％為陽性，40％為可疑陽性，游泳疲勞時(shí)間為陽性，肌電圖衰減均為陽性，與正常組小鼠之間的差異顯著；預(yù)防對(duì)照組和治療
[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.

【學(xué)位授予單位】：南京師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R392;R-332

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本文編號(hào)：1778361