胶原诱导性关节炎大鼠模型建立及其滑膜IL-33的表达

发布时间：2018-05-18 20:59

本文选题：实验性关节炎 + II型胶原　；参考：《福建医科大学》2009年硕士论文

【摘要】： 目的:优化造模条件,建立可重复的高发病率胶原诱导性关节炎(CIA)大鼠模型,并探讨胶原给药剂量以及性别因素对造模成功率的影响。检测IL-33在CIA大鼠以及正常大鼠滑膜中的表达情况并进行比较。方法:以鸡II型胶原蛋白(CII)与弗氏不完全佐剂(IFA)制成乳剂,皮内注射近交系Wistar大鼠诱导CIA。采用随机分组设计,考察400μg、300μg、200μg、0μg 4个胶原蛋白给药剂量水平,以及性别因素对发病率的影响,并每周测量足爪容积,进行关节炎评分,每两周ELISA检测血清抗CII抗体、TNF-α浓度至初次免疫后第6周,进行X线摄片并处死大鼠行病理HE染色,以检查关节病变与骨质破坏情况。选取造模成功的CIA大鼠,以半定量RT-PCR与免疫组化染色方法测定其关节滑膜IL-33的mRNA以及蛋白表达情况,并与正常对照组做比较。结果:发病大鼠可见体重下降、足掌增厚、踝关节肿胀,并有数量不等的跖、趾关节红肿,且行走不便,急性期受累足不能负重。平均临床评分达6.3±1.8,其血清抗CII抗体与TNF-α浓度均升高,初次免疫后第6周时X线及病理形态呈现慢性滑膜炎表现,可见明显骨质侵蚀。统计分析显示发病率随胶原给药剂量增加而上升,400μg组造模成功率达100%。性别对造模成功率无显著影响。各种CII剂量在造模过程中均未发生动物死亡。CIA大鼠关节滑膜IL-33 mRNA水平与蛋白表达水平均较正常对照组增高。免疫组化染色显示CIA大鼠滑膜中IL-33的主要来源有血管内皮细胞、成纤维细胞以及滑膜衬里细胞,在正常对照组大鼠滑膜的上述细胞中也可见阳性反应,但较病变滑膜浅染。结论: 1.成功建立发病稳定的大鼠模型,其临床表现显著,免疫学及病理学上具有典型不可逆慢性滑膜炎症特征,并显示出明显的骨质侵蚀。 2.提高胶原蛋白给药剂量能够提高造模成功率,400μg的剂量发病率可达100%。性别对发病率无明显影响。 3. IL-33在关节滑膜的内皮细胞、滑膜衬里细胞以及成纤维细胞中呈组成性低表达。 4.在CIA炎症情况下,滑膜IL-33表达量增加,提示其在RA等慢性炎症性疾病中可能具有重要的调控作用。
[Abstract]:Objective: to establish a rat model of CIA-induced arthritis with high incidence of collagen, and to investigate the effect of dosage of collagen and sex factors on the success rate of rat model. The expression of IL-33 in synovium of CIA rats and normal rats was detected and compared. Methods: chicken type II collagen (CII) and Freund's incomplete adjuvant (IFA) were used as emulsions. CIAs were induced by intradermal injection of inbred Wistar rats. A randomized grouping design was used to investigate the dosage of 4 collagen (400 渭 g / 300 渭 g / g) and the influence of sex factors on the incidence of the disease. The paw volume was measured weekly and the arthritis score was evaluated. The concentration of serum anti CII antibody TNF- 伪 was detected by ELISA every two weeks until the 6th week after the first immunization. X-ray films were performed and pathological HE staining was performed on the rats to examine the joint lesion and bone destruction. The expression of mRNA and protein in synovial IL-33 of CIA rats were determined by semi-quantitative RT-PCR and immunohistochemical staining, and the results were compared with those of normal control group. Results: weight loss, foot palmar thickening, ankle joint swelling, various numbers of plantaris, redness of toe joint, and inconvenient walking were observed in the rats. The average clinical score was 6.3 卤1.8, and the serum levels of anti-TNF- 伪 and anti-TNF- 伪 were increased. At the 6th week after the first immunization, the X-ray and histopathology showed chronic synovitis with obvious bone erosion. Statistical analysis showed that the incidence rate increased with the increase of the dosage of collagen, and the success rate of model making in 400 渭 g group was 100%. Sex had no significant effect on the success rate of modeling. The level of IL-33 mRNA and protein expression in synovial membrane of CII rats were higher than that of normal control group. Immunohistochemical staining showed that the main sources of IL-33 in the synovium of CIA rats were vascular endothelial cells, fibroblasts and synovial lining cells. Conclusion: 1. A stable rat model was successfully established. Its clinical manifestations were remarkable. The immunological and pathological features of chronic synovitis were typical irreversible chronic synovitis and showed obvious bone erosion. 2. Increasing the dosage of collagen can increase the success rate of model making by 400 渭 g. The incidence of collagen can reach 100 渭 g. Sex had no significant effect on morbidity. 3. IL-33 was expressed constitutively in endothelial cells, lining cells and fibroblasts of synovial membrane. 4. The increase of IL-33 expression in synovial membrane during CIA inflammation suggests that it may play an important role in the regulation of chronic inflammatory diseases such as RA.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R593.22;R-332

【共引文献】