大鼠类风湿性关节炎实验模型的建立与评价
发布时间：2019-06-12 12:48
【摘要】： 类风湿性关节炎(rheumatoid arthritis,RA)是一种以关节滑膜炎、破坏性关节病变为主要特征的慢性、进行性、侵袭性自身免疫性疾病,致畸、致残率极高且大多数患者有反复发作的病程,虽然与RA发病相关的因素已经清楚,其中,多种免疫活性细胞的活化,细胞因子、炎症介质的共同参与,血管翳的形成等是造成该病发生、发展的重要原因。但是RA的具体发病机理至今不明,尚待进一步研究。目前,改善临床症状的抗风湿药是治疗RA的主要手段,由于传统抗风湿药的毒副作用较大,而生物制剂价格昂贵,因而患者长期服用依从性差,不利于疾病控制。因此研制开发有效、毒副作用小的抗风湿药物势在必行。而研究并建立合理有效的类风湿性关节炎动物模型是研制开发有效治疗药物和制订有效治疗措施的关键。 本研究以Wistar大鼠为试验动物,用不同的实验试剂对大鼠进行免疫,从关节炎指数评定、血清学检测、病理学检测和影像学检测等方面对模型进行综合评价。方法是:将4～6周龄雌性Wistar大鼠随机分成五组:M1组(CFA足跖部皮下注射法免疫)、M2组(鸡CII+CFA乳化剂足跖部、尾根部、背部三点注射法免疫)、M3组(鸡CII+CFA乳化剂+冰浴7d)、M4组(鸡CII蛋白免疫)、对照组(不做任何处理),7d后足跖部皮下注射加强免疫一次。于大鼠首次免疫后7d、14d、21d分别测量各组大鼠的足跖部厚度,并进行关节炎指数(AI)评定;于21d、42d采用双抗体夹心ELISA法测定各组大鼠血清中抗CII抗体水平、细胞因子TNF-α、IL-1β和IL-17水平,并于21d和42d分别观察大鼠的放射学变化和组织病理学变化。通过以上几方面指标的检测综合评价建立的大鼠实验性关节炎模型。 研究结果表明:M1～M3组大鼠在首次免疫10d后不同的测定时间点AI值和足跖部厚度均显著高于对照组和M4组大鼠(P＜0.05),炎症高峰期出现于首次免疫后的19～27d,而M4组与对照组的大鼠比较不具有统计学意义。M2、M3组大鼠血清中抗CII抗体水平在炎症高峰期时极显著的高于对照组(P＜0.01),而M1组和M4组大鼠血清中抗CII抗体水平则无明显变化。M1～M3组大鼠在炎症高峰期时血清中TNF-α、IL-1β、IL-17水平均显著高于对照组大鼠(P＜0.05),但高峰期后炎症水平逐渐有所下降,但42d仍高于正常水平。病理组织切片显示,M1～M3组大鼠在关节滑膜中有大量炎性细胞浸润,骨细胞增生,并形成血管翳。X光片显示踝关节周围软组织严重肿胀,踝关节、足趾关节间隙模糊、狭窄,骨质破坏。 通过以上指标的综合比较,给予不同免疫剂的4个试验组大鼠中,鸡CII+CFA乳化剂+冰浴(M3)组大鼠无论从外观表现、血清学检测指标还是从组织病理学、放射学等方面指标均与人类RA极为相似,且比其它模型组大鼠更符合人类RA的特点,这为进一步深入研究人类RA的发病机制及新药的研发奠定了坚实的基础。
[Abstract]:Rheumatoid arthritis (rheumatoid arthritis,RA) is a chronic, progressive, invasive autoimmune disease characterized by joint synovitis and destructive joint disease, teratogenic, disability rate is extremely high and most patients have recurrent disease course, although the factors related to the pathogenesis of RA are already clear, in which the activation of a variety of immunoactive cells, cytokines and inflammatory mediators are involved. The formation of pannus is an important cause of the occurrence and development of the disease. However, the specific pathogenesis of RA is still unknown and needs to be further studied. At present, antirheumatic drugs to improve clinical symptoms are the main means to treat RA. Because of the large toxic and side effects of traditional antirheumatic drugs and the high price of biological agents, the long-term compliance of patients is poor, which is not conducive to disease control. Therefore, it is imperative to develop effective antirheumatic drugs with little toxic and side effects. The research and establishment of a reasonable and effective animal model of rheumatoid arthritis is the key to the development of effective therapeutic drugs and effective treatment measures. In this study, Wistar rats were immunized with different experimental reagents, and the model was comprehensively evaluated from the aspects of arthritis index evaluation, serological detection, pathological detection and imaging detection. Methods: female Wistar rats aged 4 to 6 weeks were randomly divided into five groups: M1 group (CFA foot metatarsal injection method), M2 group (chicken CII CFA emulsifier foot metatarsal, tail root, back three-point injection), M3 group (chicken CII CFA emulsifier ice bath 7 days), M4 group (chicken CII protein immunization), control group (no treatment), 7 days later, foot metatarsal subcutaneous injection enhanced immunization. The metatarsal thickness of each group was measured on the 7th day, 14th day and 21st day after the first immunization, and the arthritis index (AI) was used to evaluate the arthritis index. On the 21st day, the levels of anti-CII antibody, cytokines TNF- 伪, IL-1 尾 and IL-17 in serum were measured by double antibody sandwich ELISA method, and the radiological and histopathological changes were observed on the 21st day and 42nd day, respectively. The experimental arthritis model of rats was established by comprehensive evaluation of the above indexes. The results showed that the AI value and foot metatarsal thickness in M1~M3 group were significantly higher than those in control group and M4 group at different time points after the first immunization. The peak period of inflammation occurred at 19 鈮，
本文编号：2498039
[Abstract]:Rheumatoid arthritis (rheumatoid arthritis,RA) is a chronic, progressive, invasive autoimmune disease characterized by joint synovitis and destructive joint disease, teratogenic, disability rate is extremely high and most patients have recurrent disease course, although the factors related to the pathogenesis of RA are already clear, in which the activation of a variety of immunoactive cells, cytokines and inflammatory mediators are involved. The formation of pannus is an important cause of the occurrence and development of the disease. However, the specific pathogenesis of RA is still unknown and needs to be further studied. At present, antirheumatic drugs to improve clinical symptoms are the main means to treat RA. Because of the large toxic and side effects of traditional antirheumatic drugs and the high price of biological agents, the long-term compliance of patients is poor, which is not conducive to disease control. Therefore, it is imperative to develop effective antirheumatic drugs with little toxic and side effects. The research and establishment of a reasonable and effective animal model of rheumatoid arthritis is the key to the development of effective therapeutic drugs and effective treatment measures. In this study, Wistar rats were immunized with different experimental reagents, and the model was comprehensively evaluated from the aspects of arthritis index evaluation, serological detection, pathological detection and imaging detection. Methods: female Wistar rats aged 4 to 6 weeks were randomly divided into five groups: M1 group (CFA foot metatarsal injection method), M2 group (chicken CII CFA emulsifier foot metatarsal, tail root, back three-point injection), M3 group (chicken CII CFA emulsifier ice bath 7 days), M4 group (chicken CII protein immunization), control group (no treatment), 7 days later, foot metatarsal subcutaneous injection enhanced immunization. The metatarsal thickness of each group was measured on the 7th day, 14th day and 21st day after the first immunization, and the arthritis index (AI) was used to evaluate the arthritis index. On the 21st day, the levels of anti-CII antibody, cytokines TNF- 伪, IL-1 尾 and IL-17 in serum were measured by double antibody sandwich ELISA method, and the radiological and histopathological changes were observed on the 21st day and 42nd day, respectively. The experimental arthritis model of rats was established by comprehensive evaluation of the above indexes. The results showed that the AI value and foot metatarsal thickness in M1~M3 group were significantly higher than those in control group and M4 group at different time points after the first immunization. The peak period of inflammation occurred at 19 鈮，
本文编号：2498039
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