减毒百日咳杆菌注射部位与实验性变态反应性脑脊髓炎大鼠模型诱导效果的关系(英文)

Relation between the injected site of pertussis toxin and the induced effect on experimental allergic encephalomyelitis in rats

暂未订购

导出

摘要背景:国外实验室一般选用近交系Lewis大鼠诱导实验性变态反应性脑脊髓炎模型,但国内缺乏Lewis大鼠。因此,本实验选用了国内数量充足、但不十分敏感的Wistar大鼠,在加用百日咳减毒活菌的情况下,成功地在Wistar大鼠中诱导出实验性变态反应性脑脊髓炎动物模型。目的:探讨在不同部位加用减毒百日咳杆菌对实验性变态反应性脑脊髓炎非敏感品系Wistar大鼠模型诱导的影响。设计:随机对照动物实验。单位:山西大同大学脑科学研究所。材料:实验于2003-03/10在山西大同大学脑科学研究所完成。雌性Wistar大鼠58只随机分为3组:①足背实验性变态反应性脑脊髓炎组24只。②腹腔实验性变态反应性脑脊髓炎组24只。③佐剂组10只。方法:除常规免疫动物外,足背实验性变态反应性脑脊髓炎组大鼠于足背皮内或皮下注射减毒百日咳杆菌0.05mL/只(含5.0×1010个菌体);腹腔实验性变态反应性脑脊髓炎组大鼠于腹腔注射减毒百日咳杆菌0.05mL/只(含5.0×1010个菌体);佐剂组则以完全弗氏佐剂代替抗原。主要观察指标:①发病时间。②体质量变化。③临床症状。④脑组织病理情况。结果:58只大鼠均进入结果分析。①发病率和发病时间:足背实验性变态反应性脑脊髓炎组发病率为87.5%(21/24),发病时间为免疫后(10.25±1.67)d,而腹腔实验性变态反应性脑脊髓炎组分别为35.7%(9/24)和(14.8±1.79)d,两组比较差异有显著性意义(P<0.05)。②体质量变化和临床症状:足背实验性变态反应性脑脊髓炎组体质量变化为(-16.00±7.30)g,症状评分为3.4±0.7;腹腔实验性变态反应性脑脊髓炎组体质量变化为(-9.14±13.11)g,症状评分为2.4±0.5。③脑组织病理情况:佐剂组大鼠脑脊髓组织没有或有少数单个核细胞浸润。实验性变态反应性脑脊髓炎大鼠炎症病灶侵犯脊髓腰膨大的白质以及灰白质交界处、软脊膜及脊髓实质,大脑皮质及皮髓质交界处甚至深部髓质、脑脊膜和侧脑室周围也被侵犯。病灶也累及小脑、脑干和视交叉,这与观察到的共济障碍、抽搐等表现相一致。苏木精-伊红染色显示病变血管周围有淋巴细胞和单核细胞浸润,呈典型的袖套样改变。足背实验性变态反应性脑脊髓炎和腹腔实验性变态反应性脑脊髓炎发病大鼠均有典型的袖套样改变,但足背实验性变态反应性脑脊髓炎大鼠病灶多。结论:足背皮下注射百日咳毒素诱导实验性变态反应性脑脊髓炎模型病程、病理改变、临床表现都很典型,且发病率高,经济易行,是一种较为理想的实验性变态反应性脑脊髓炎模型诱导方法。 BACKGROUND： The animals commonly used to induce experimental allergic encephalomyelitis （EAE） in oversea laboratory are rodentia animals such as Lewis rats. But in China we are short of Lewis rats. The un-susceptire animal Wistar rats arc inexpensive and plentiful. The adding of pertussis toxin may induce EAE successfully in EAE un-susceptive Wistar rats. OBJECTIVE： To investigate the influence of pertussis toxin injected at different sites in inducing EAE model in un-susceptive Wistar rats. DESIGN： A randomized control animal experiment. SETTING： Institute of Brain Science, shanxi Datong University. MATERIALS： The study was performed in the Institute of Brain Science of Shanxi Datong University from March to October in 2003. Fifty-eight female Wistar rats were randomly divided into three groups：（1） foot dorsum EAE group （n=24）; （2） intraperitoneal EAE group （n=24）; （3） complete Freund adjuvant （CFA） group （n=10）. METHODS： Besides routine immunization, each rat.in the foot dorsum EAE group and intraperitoneal EAE group was administrated with 0.05 mL pertussis toxin （containing 5.0×10^10 thalli）, which were given intraperitoneally and subcutaneously on one hind foot respectively, and the antigen in the CFA group was replaced by CFA. MAIN OUTCOME MEASURES：（1） Time of attack; （2） change of body mass; （3） clinical symptoms; （4） pathological conditions of brain tissue. RESULTS： All the 58 rats were involved in the analysis of results. （1） Incidence rate and time of attack： In the foot dorsum EAE group, the incidence of EAE was 87.5% （21/24）, and the time of attack was at （10.25 ±1.67） days after immunization, which were significantly different from those in .theintraperitoneal EAE group [35.7% （9/24）, （14.8±1.79） days, P 〈 0.05]. （2） Change of body mass and clinical symptoms： In the foot dotsum EAE group, the change of body mass was （-16.00±7.30） g and the symptom score was 3.4±0.7, and those in the intraperitoneal EAE group were [（-9.14± 13.11） g, 2.4±0.5]. （3） Pathological conditions of brain tissue： There were no or little infiltration of inflammatory cells in the encephalon and spinal cord of CFA rats. In the EAE rats, there were inflammatory cells infiltrated in the boundary of white matter and gray matter of lumbar intumescence, spinal pie mater, spinal parenchyma, and the boundary of cerebral cortex and medulla, even deep medulla, meninges and around lateral ventricle. There were also mild inflammations in the cerebellum, brainstem and optic chiasma, which were concordant with the observed asynchronism, tic, etc. Hematoxylin and eosin （HE） staining displayed that the infiltrated mononuclear cells assembled in perivascular spaces, which were identified by morphological criteria as lymphocyte and macrophages. Forming typical muff-like changes, the inflammation was less severe in intraperitoneal EAE group than in subcutaneous foot dorsum EAE group. CONCLUSION： The EAE model induced in Wistar rats by Pertussis toxin administered subcutaneously on foot dorsum has the. representative course of diseases, pathology change and clinical manifestation and the incidence of diseases is high and the cost is low. So it is a more ideal EAE modal inducing method.