卵清蛋白诱导的兔类风湿关节炎模型的建立及评估

Establishment and evaluation of rabbit rheumatoid arthritis model induced by ovalbumin

目的建立并评估卵清蛋白(OVA)诱导的兔类风湿关节炎(RA)模型。方法于家兔肩胛间区的5个部位进行皮下注射OVA,每周1次,连续3 w,对RA组进行致敏,注射2 w后,在兔双侧关节腔内注射OVA建立RA模型。对照组以同样方法注射等量的生理盐水。建模后每周分别用千分游标卡尺和电子体温计测量关节直径和关节表面皮肤温度;第1、4、8周采用ELISA试剂盒检测各组家兔外周血清中的IL-1、TNF-α和抗OVA IgG水平;于建模后第1、4、8周处死家兔,取腹股沟淋巴结,用不同浓度OVA刺激淋巴细胞增殖,检测淋巴细胞对特异性抗原的反应;HE染色观察膝关节滑膜增生、软骨破坏程度及缺损周围组织炎性细胞浸润情况。结果 RA组家兔关节腔注射OVA后1 w,关节直径明显增大,表面皮肤温度明显升高,随后逐渐降低至稳定的水平,但仍明显高于对照组;在关节腔注射抗原后第1、4、8周,RA组血清中IL-1、TNF-α和抗OVA IgG水平较对照组均显著升高(P<0.01);RA组淋巴细胞增殖实验与对照组相比,差异有统计学意义(P<0.05);RA组家兔关节滑膜大量增生,第8周时软骨破坏明显;组织切片可见滑膜组织大量增生,炎症细胞浸润,软骨层破坏。结论 OVA诱导的兔RA模型能够很好地模拟人RA病理过程,可成为研究RA的理想的动物模型。

Objective To establish and evaluate rabbit rheumatoid arthritis （RA） model induced by ovalbumin （OVA）. Methods After continuous subcutaneous injection of OVA at 5 points of scapular area each week to sensitize RA group rabbits for three weeks, 2 weeks later after sensitization, OVA was injected into the intra-articular of each knee to induce RA models. And control group were given the same amount of saline in the same way. The joint diameter and surface skin temperature changed with micrometer caliper and electronic clini- cal thermometer weekly; IL-1, TNF-a and anti-OVA IgG levels in the serum were determined with ELISA Kits at first, fourth and eighth week. Rabbits were sacrificed at the second ,fourth and eighth week after articular injection to obtain inguinal lymph nodes stimulated by dif- ferent concentrations of OVA for lymphocyte proliferation test to detect lymphocyte responses to specific antigen. The knees and synovial tis- sue were removed to observe synovial hyperplasia , cartilage destruction and inflammatory cells infiltration by HE staining. Results The joint diameter and surface skin temperature of RA group increased significantly after articular injection for a week, then gradually reduced to a sta- ble level, but still significantly higher than those of control group. The IL-1, TNF-c^and anti-OVA IgG levels in the serum of RA group were significantly higher than those of control group at the first ,fourth and eighth week （P 〈 0. 01 ） ; Lymphocyte proliferation test also indicated statistically significant difference （ P 〈 0. 05 ） between the two groups. Macroscopic observation showed obvious synovial hyperplasia and car- tilage destruction saerificed at eighth week. Tissue sections showed synovial tissue hyperplasia, inflammatory cells infiltration and cartilage destruction. Conclusions RA model induced by OVA can imitate the pathological process of human RA very well, offering an ideal animal model of RA.