【摘要】： 類風濕性關節(jié)炎(rheumatoid arthritis,RA)是一種以關節(jié)滑膜炎、破壞性關節(jié)病變?yōu)橹饕卣鞯穆�、進行性、侵襲性自身免疫性疾病,致畸、致殘率極高且大多數(shù)患者有反復發(fā)作的病程,雖然與RA發(fā)病相關的因素已經清楚,其中,多種免疫活性細胞的活化,細胞因子、炎癥介質的共同參與,血管翳的形成等是造成該病發(fā)生、發(fā)展的重要原因。但是RA的具體發(fā)病機理至今不明,尚待進一步研究。目前,改善臨床癥狀的抗風濕藥是治療RA的主要手段,由于傳統(tǒng)抗風濕藥的毒副作用較大,而生物制劑價格昂貴,因而患者長期服用依從性差,不利于疾病控制。因此研制開發(fā)有效、毒副作用小的抗風濕藥物勢在必行。而研究并建立合理有效的類風濕性關節(jié)炎動物模型是研制開發(fā)有效治療藥物和制訂有效治療措施的關鍵。 本研究以Wistar大鼠為試驗動物,用不同的實驗試劑對大鼠進行免疫,從關節(jié)炎指數(shù)評定、血清學檢測、病理學檢測和影像學檢測等方面對模型進行綜合評價。方法是:將4～6周齡雌性Wistar大鼠隨機分成五組:M1組(CFA足跖部皮下注射法免疫)、M2組(雞CII+CFA乳化劑足跖部、尾根部、背部三點注射法免疫)、M3組(雞CII+CFA乳化劑+冰浴7d)、M4組(雞CII蛋白免疫)、對照組(不做任何處理),7d后足跖部皮下注射加強免疫一次。于大鼠首次免疫后7d、14d、21d分別測量各組大鼠的足跖部厚度,并進行關節(jié)炎指數(shù)(AI)評定;于21d、42d采用雙抗體夾心ELISA法測定各組大鼠血清中抗CII抗體水平、細胞因子TNF-α、IL-1β和IL-17水平,并于21d和42d分別觀察大鼠的放射學變化和組織病理學變化。通過以上幾方面指標的檢測綜合評價建立的大鼠實驗性關節(jié)炎模型。 研究結果表明:M1～M3組大鼠在首次免疫10d后不同的測定時間點AI值和足跖部厚度均顯著高于對照組和M4組大鼠(P＜0.05),炎癥高峰期出現(xiàn)于首次免疫后的19～27d,而M4組與對照組的大鼠比較不具有統(tǒng)計學意義。M2、M3組大鼠血清中抗CII抗體水平在炎癥高峰期時極顯著的高于對照組(P＜0.01),而M1組和M4組大鼠血清中抗CII抗體水平則無明顯變化。M1～M3組大鼠在炎癥高峰期時血清中TNF-α、IL-1β、IL-17水平均顯著高于對照組大鼠(P＜0.05),但高峰期后炎癥水平逐漸有所下降,但42d仍高于正常水平。病理組織切片顯示,M1～M3組大鼠在關節(jié)滑膜中有大量炎性細胞浸潤,骨細胞增生,并形成血管翳。X光片顯示踝關節(jié)周圍軟組織嚴重腫脹,踝關節(jié)、足趾關節(jié)間隙模糊、狹窄,骨質破壞。 通過以上指標的綜合比較,給予不同免疫劑的4個試驗組大鼠中,雞CII+CFA乳化劑+冰浴(M3)組大鼠無論從外觀表現(xiàn)、血清學檢測指標還是從組織病理學、放射學等方面指標均與人類RA極為相似,且比其它模型組大鼠更符合人類RA的特點,這為進一步深入研究人類RA的發(fā)病機制及新藥的研發(fā)奠定了堅實的基礎。
[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- 偽, 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，

本文編號：2498039