發(fā)布時間：2018-02-03 11:21

  本文關(guān)鍵詞： 肌肉特異性激酶 重癥肌無力 單克隆抗體 免疫熒光 透射電鏡　出處：《延邊大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的：通過建立Lewis大鼠實驗性自身免疫性重癥肌無力(myasthenia gravis,MG)模型(experimental autoimmune MG, EAMG),以此研究肌肉特異性激酶(muscle-specific kinase, MuSK)抗體(muscle-specific kinase antibody, MuSKAb)和MG發(fā)病機(jī)制的關(guān)系。 方法：選取10只雌性Lewis大鼠,隨機(jī)分為實驗組和對照組。采用被動免疫的方法,將MuSK單克隆抗體通過腹腔注射的方式給予實驗組大鼠,對照組大鼠采用同樣的方法給藥,每2小時觀察1次大鼠的飲食飲水情況以及有無臨床癥狀出現(xiàn),并隨時記錄大鼠的體重變化。給藥96小時后,迅速處死兩組大鼠,并取下大鼠的肋間肌、腓腸肌和脛前肌,與-80℃冰箱中保存。之后對于取下的肌肉組織進(jìn)行免疫熒光實驗,用以觀察抗MuSK單克隆抗體和MuSK蛋白結(jié)合情況和C3補(bǔ)體激活情況。同時做透射電鏡實驗,用以觀察神經(jīng)肌肉接頭處(NMJ)的形態(tài)學(xué)變化。 結(jié)果：在給藥96小時后實驗組與對照組相比,未出現(xiàn)抓握和嘶叫無力的明顯臨床癥狀。經(jīng)過免疫熒光實驗后發(fā)現(xiàn),實驗組肋間肌、腓腸肌和脛前肌的細(xì)胞膜表面出現(xiàn)明顯的黃綠色熒光和紅色熒光物質(zhì),黃綠色熒光說明MuSK單克隆抗體和跨膜蛋白MuSK發(fā)生特異性結(jié)合,紅色熒光說明C3補(bǔ)體抗體和C3發(fā)生特異性結(jié)合。而對照組無熒光出現(xiàn)。透射電鏡觀察顯示實驗組大鼠運(yùn)動終板突觸后區(qū)域簡單化,突觸后膜皺褶退化,分級減少并短縮,突觸間隙略增寬。而實驗組無明顯變化。 結(jié)論：將MuSK單克隆抗體注射給大鼠后未能引起肌無力的臨床癥狀,但在細(xì)胞膜處發(fā)現(xiàn),MuSKAb和MuSK發(fā)生特異性結(jié)合,并且能夠激活C3補(bǔ)體,同時NMJ發(fā)生了超微結(jié)構(gòu)形態(tài)學(xué)變化。抗MuSKAb的致病性仍需進(jìn)一步研究。
[Abstract]:Objective: to establish myasthenia gravis of experimental autoimmune myasthenia gravis in Lewis rats. Experimental autoimmune (EAMG). To study muscle-specific kinase. The relationship between muscle-specific kinase antibody (MuSKAb) and the pathogenesis of MG. Methods: ten female Lewis rats were randomly divided into experimental group and control group. The MuSK monoclonal antibody was injected intraperitoneally into the experimental group by passive immunization. Rats in the control group were treated with the same method. The diet and drinking water status and clinical symptoms were observed once every 2 hours, and the weight changes of the rats were recorded at any time. 96 hours after administration of the drug. The intercostal muscle, gastrocnemius muscle and tibial anterior muscle of the rats were quickly killed and stored in refrigerator at -80 鈩，

本文編號：1487248