柯萨奇病毒B诱导的免疫性多发性肌炎模型的建立

Preliminary Study on the Establishment of an Animal Model of Experimental Autoimmune Polymyositis Infution with Coxsaekievirus B

目的：探讨制作实验性免疫性多发性肌炎动物模型的方法,以及柯萨奇病毒感染与多发性肌炎发病的关系。方法：分别用不同毒力0．3ml的柯萨奇病毒B_1(CVB_1 10^(-2),10^(-3),10^(-4) TCID_(50))感染和兔肌肉匀浆(30mg/ml)加完全弗氏佐剂多次免疫28只正常豚鼠,共观察3～5周；和不同毒力0．2mlCVB_1 10^(-3),10^(-5),10^(-7)TCID_(50)分别腹腔接种各20只BALB/C和C57BL/6小鼠；以及0．1ml CVB_3 10^(-6)TCID_(50)腹腔接种28只BALB/C小鼠,共观察1～3周,制成实验性多发性肌炎动物模型,观察其在血清肌酶、巨检和组织病理的改变。结果：发现0．3ml毒力为10^(-2)(A组),10^(-3)(B组)和10^(-4)(C组)TCID_(50)柯萨奇病毒B_1感染豚鼠组,3周后仅少数豚鼠出现多发性肌炎的症状,但均能导致肌酶谱(CK,CK-MM,LDH和AST)异常升高,与正常组相比有明显差异(均P＜0．05),以B组升高值最高,但巨检肌肉无异常；4周后10^(-3)组(F组)豚鼠33．3%出现下肢瘫痪,巨检发现肌肉萎缩,弹性差,病理检查证实为多发性肌炎改变；3周与5周的CK和CK-MM升高值相比,无显著差异(P=0．1031和P=0．1164)。单纯兔肌肉匀浆免疫(E组)也能导致豚鼠肌酶谱异常升高,但不如病毒感染后明显,也不引起股肌肉肌炎的病理改变。柯萨奇病毒B_1和B_3均不能诱导BALB/C和C57BL/6小鼠出现多发性肌炎改变。结论：柯萨奇病毒B_1感染的毒力和病毒量与豚鼠多发性肌炎的发病相关,在感染5周后比较明显。本模型可作为研究人类多发性肌炎的一个重要手段,为人类肌炎的发病机理和治疗提供较好的动物模型。

Objective：To investigate the methods of the establishment of an animal model of experimental autoimmune polymyosiris, and to understand the relation between infection with eox.sackievirus B （CVB） and onset of polymyositis in the models of animals. Methods：28 guinea pigs were innoculated intraperitoneally with 0.3 ml of 10^-2, 10^-3 and 10^-4 TCID50 CVB1 in groups A,B,C and F separately, and immunized with 6mg rabbit muscle homogenates in complete freund＇s adjuvant（CFA） once a week, in group D only with it for 3-5 weeks. Both 20 Balb/c and C57bl/6 mice were intraperitoneally innoculated with 0. 2ml of CVB1 10^-3,10^-5 and 10^-7 TCID50 for 1 -3 weeks separately. 28 Balb/c mice were intraperitoneally innoculated with 0. lml of CVB3 10^-6 TCID50 for 1-3 weeks. Serum enzymes of CK,CK-MM,LDH and AST, gross examination and thigh muscule histopathological changes were observed. Resdts： Few guinea pigs infected with 0. 3ml CVB1 of toxicity 10^-2 （group A）, 10^-3 （group B）and 10^-4 （group C）TCID50 showed typical manifestations of polymyositis in 3 weeks after viral infection, and that the serum enzymes （CK,CK-MM,LDH and AST） evidently elevated than the normal control（P 〈0. 05）. But the gross examinations and the thigh muscle histopathological changes were normal. And in group E, although it could cause the elevation of the serum enzymes evidently, but the value was lower than other groups of A,B and C. All Balb/c and C57BL/6 mice infected with 0. 2ml CVB1 of toxicity 10^-3 , 10^-5 and 10 7 TCID50 could not induce the polymyositis （both in serum enzymes, gross examinations and histopatbological changes were normal）. All Balb/e mice infected with 0. 1ml CVB3 10^-6 TCID50 couled not be induced with the polymyositis, although it could induce viral myocarditis. Conclusion： Not only infection with coxsackievirus B1 could induce PM, but also its quantity and toxicity were correlated with the onset of polymyositis. It was evident in 5 weeks after viral infection. This method of establishing an animal model of experimental autoimmune polymyositis were feasible,dependable and successful.