摘要
目的从丁氏双鳍电鳐的电器官分离乙酰胆碱受体(AChR)免疫大鼠,建立实验性自身免疫性重症肌无力(EAMG)动物模型。方法参照徐氏法从丁氏双鳍电鳐的电器官提取AChR蛋白,并采用Folin-酚试剂法及酶联免疫吸附法检测提取蛋白的含量及活性;以提取的蛋白主动免疫Lewis大鼠,每天观察其临床表现,并于免疫后7周进行低频重复电刺激、单纤维肌电图及血清中AChRAb水平的检测。结果提取蛋白的含量为35.4mg/mL,AChR提取物可以与阳性血清中的AChRAb发生结合反应。免疫后1周左右,实验组有3只大鼠出现轻度的肌无力症状,持续2￣5d自行缓解;免疫后5周左右实验组8只大鼠均表现出不同程度的肌无力症状。实验组大鼠低频重复电刺激阳性率达75%,其衰减率明显高于对照组。实验组大鼠单肌纤维动作电位平均连续差(MCD)值与正常对照组比较明显延长(P<0.01),而福氏完全佐剂(CFA)对照组与正常对照组间MCD值无明显差异。实验组大鼠AChRAb水平明显高于CFA对照组(P<0.01);实验组7只大鼠AChRAb的P/N值>2.1,而CFA对照组P/N值均<1.4。结论从丁氏双鳍电鳐的电器官提取的AChR蛋白成功地诱导EAMG动物模型,可为重症肌无力进一步研究创造条件。
[Objective] To study the animal model of experimental autoimmune myasthenia gravis (EAMG) induced with extract from electric organ of Narcine timilei in rats. [Methods] Lewis rats were immunized with the protein extracted from electric organ of Narcine timilei, the clinical manifestations were observed every day, and repetitive nerve stimulation (RNS),single fiber electromyography (SFEMG) and the levels of acetylcholine receptor antibody (AChRAb) were detected on the 49th days post immunization. [Results] The content of the protein extracted from electric organ of Narcine timile was 35.4 mg/mL, and the protein could respond with AChRAb from the serum of seropositive myasthenia gravis (MG) patient. The mild symptoms were observed on 3 experimental rats after a week, which lasted for 2 to 5 days. And after 5 weeks, all experimental rats presented some symptoms at different degree like MG. The positive rate of RNS of the experimental group was 75%, and the attenuation rate of these rats was higher than that of control rats. The mean consecutive difference (MCD) of experimental rats were longer than that of control rats markedly (P＜0.01). The levels of AChRAb of experimental rats were higher than that of CFA control rats significantly (P＜0.01), and the values of P/N of 7 experimental rats were more than 2.1, those of the control rats were less than 1.4. [Conclusion] These pointed out that the extract from electric organ of Narcine timilei could serve as the antigen to induce EAMG for further research on MG.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2006年第24期3681-3684,共4页
China Journal of Modern Medicine
关键词
重症肌无力
乙酰胆碱受体
主动免疫
电鳐
myasthenia gravis
acetylcholine receptor
active immunity
Narcine timilei
