本文選題：實驗性關(guān)節(jié)炎 + II型膠原�。� 參考：《福建醫(yī)科大學》2009年碩士論文

【摘要】： 目的:優(yōu)化造模條件,建立可重復的高發(fā)病率膠原誘導性關(guān)節(jié)炎(CIA)大鼠模型,并探討膠原給藥劑量以及性別因素對造模成功率的影響。檢測IL-33在CIA大鼠以及正常大鼠滑膜中的表達情況并進行比較。 方法:以雞II型膠原蛋白(CII)與弗氏不完全佐劑(IFA)制成乳劑,皮內(nèi)注射近交系Wistar大鼠誘導CIA。采用隨機分組設計,考察400μg、300μg、200μg、0μg 4個膠原蛋白給藥劑量水平,以及性別因素對發(fā)病率的影響,并每周測量足爪容積,進行關(guān)節(jié)炎評分,每兩周ELISA檢測血清抗CII抗體、TNF-α濃度至初次免疫后第6周,進行X線攝片并處死大鼠行病理HE染色,以檢查關(guān)節(jié)病變與骨質(zhì)破壞情況。選取造模成功的CIA大鼠,以半定量RT-PCR與免疫組化染色方法測定其關(guān)節(jié)滑膜IL-33的mRNA以及蛋白表達情況,并與正常對照組做比較。 結(jié)果:發(fā)病大鼠可見體重下降、足掌增厚、踝關(guān)節(jié)腫脹,并有數(shù)量不等的跖、趾關(guān)節(jié)紅腫,且行走不便,急性期受累足不能負重。平均臨床評分達6.3±1.8,其血清抗CII抗體與TNF-α濃度均升高,初次免疫后第6周時X線及病理形態(tài)呈現(xiàn)慢性滑膜炎表現(xiàn),可見明顯骨質(zhì)侵蝕。統(tǒng)計分析顯示發(fā)病率隨膠原給藥劑量增加而上升,400μg組造模成功率達100%。性別對造模成功率無顯著影響。各種CII劑量在造模過程中均未發(fā)生動物死亡。CIA大鼠關(guān)節(jié)滑膜IL-33 mRNA水平與蛋白表達水平均較正常對照組增高。免疫組化染色顯示CIA大鼠滑膜中IL-33的主要來源有血管內(nèi)皮細胞、成纖維細胞以及滑膜襯里細胞,在正常對照組大鼠滑膜的上述細胞中也可見陽性反應,但較病變滑膜淺染。 結(jié)論: 1.成功建立發(fā)病穩(wěn)定的大鼠模型,其臨床表現(xiàn)顯著,免疫學及病理學上具有典型不可逆慢性滑膜炎癥特征,并顯示出明顯的骨質(zhì)侵蝕。 2.提高膠原蛋白給藥劑量能夠提高造模成功率,400μg的劑量發(fā)病率可達100%。性別對發(fā)病率無明顯影響。 3. IL-33在關(guān)節(jié)滑膜的內(nèi)皮細胞、滑膜襯里細胞以及成纖維細胞中呈組成性低表達。 4.在CIA炎癥情況下,滑膜IL-33表達量增加,提示其在RA等慢性炎癥性疾病中可能具有重要的調(diào)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- 偽 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 鹵1.8, and the serum levels of anti-TNF- 偽 and anti-TNF- 偽 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.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R593.22;R-332

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