本文選題：自身免疫性炎性肌病 + 動物模型��； 參考：《北京協(xié)和醫(yī)學院》2011年博士論文

【摘要】：[目的]1.完善自身免疫性炎性肌病大鼠模型的制作；2.探討自身免疫性炎性肌病Lewis大鼠模型發(fā)生肺間質(zhì)病變的情況,建立穩(wěn)定的自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型；3.探索自身免疫性炎性肌病Lewis大鼠模型肺間質(zhì)病變的發(fā)生機制；4.探討TRAIL及其受體在自身免疫性炎性肌病Lewis大鼠模型肺間質(zhì)病變的表達和作用：5.探索糖皮質(zhì)激素對自身免疫性炎性肌病Lewis大鼠模型肺間質(zhì)病變的影響及其機制。 [方法]1.將SD大鼠、Westar大鼠和Lewis大鼠根據(jù)品系、性別、免疫物劑量等因素分組,通過兔骨骼肌勻漿等比例混合弗氏完全佐劑多點皮下注射免疫的方法建立自身免疫性炎性肌病大鼠模型,取股四頭肌進行組織病理學檢查,比較各組之間成模率、死亡率、肌肉病理評分等觀察指標的差異；2.通過兔骨骼肌勻漿等比例混合弗氏完全佐劑多點皮下注射免疫的方法建立自身免疫性炎性肌病Lewis大鼠模型,取雙側(cè)肺行組織病理學檢查和免疫組化檢測,觀察其肺部病變；3.通過免疫組化染色結(jié)合計算機圖像分析方法檢測自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺組織中TGF-β、CTGF、ICAM、VCAM及CD163等分子的表達水平,比較與對照組之間的差別；4.通過免疫組化染色結(jié)合計算機圖像分析方法檢測自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺組織中TRAIL及其受體的表達,并分析其與肺組織病理改變之間的相關性；5.比較不同劑量、不同治療起點對自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺纖維化評分以及局部肺組織中TGF-β、CTGF、ICAM、VCAM等分子表達水平的影響。 [結(jié)果]1.三個大鼠品系中,Lewis大鼠模型成功率最高,且實驗耐受性最好,相同免疫劑量下雌性組的成模率均高于雄性組,注射兔骨骼肌勻漿蛋白劑量從8mg/kg增加至1 Omg/kg時,成模率及模型病理評分明顯升高,但從1 Omg/kg增加至12mg/kg時,成模率及模型病理評分無明顯變化,而大鼠死亡率升高,免疫4～5周時可見骨骼肌纖維壞死、非壞死肌纖維及間質(zhì)血管周圍炎細胞浸潤等典型病變；2.部分自身免疫性炎性肌病模型組Lewis大鼠肺組織出現(xiàn)不同程度肺間質(zhì)病變表現(xiàn),發(fā)生率為68.8%。典型的間質(zhì)病變?yōu)殚g質(zhì)性肺泡炎性反應和肺間質(zhì)纖維化改變,最多見的病理類型為非特異性間質(zhì)性肺炎(NSIP)和尋常型間質(zhì)性肺炎(UIP),免疫組織化學染色發(fā)現(xiàn)增厚肺泡間隔內(nèi)可見大量CD8+T淋巴細胞,支氣管及血管周圍可見大量CD20+B淋巴細胞；3.免疫組化染色結(jié)合計算機圖像分析結(jié)果顯示自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺組織中TGF-β和CTGF的表達量明顯高于對照組,特別是血管和支氣管周圍的陽性反應最為明顯。同時ICAM和VCAM的表達量明顯高于對照組。CD163陽性細胞數(shù)明顯高于對照組。4.免疫組化染色結(jié)合計算機圖像分析方法顯示自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺組織中TRAIL表達明顯升高、DCR1表達明顯降低。同時肺間質(zhì)病變中浸潤淋巴細胞上部分高表達DCR1。相關性分析發(fā)現(xiàn)肺泡炎性反應程度與肺組織中CD8+T淋巴細胞的浸潤有關,肺纖維化程度與肺組織中TRAIL的表達升高有關；5.大劑量糖皮質(zhì)激素可有效減輕自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺間質(zhì)纖維化程度,小劑量、早干預則效果更優(yōu),其機制可能與減少病變局部TGF-β、CTGF和ICAM等炎癥和促纖維化因子以及凋亡相關因子TRAIL和DR4的表達有關。 [結(jié)論]1.在應用異種動物骨骼肌勻漿多點皮下注方法建立自身免疫性炎性肌病大鼠模型時,選擇雌性Lewis大鼠作為實驗對象,給予兔骨骼肌勻漿1 Omg/kg連續(xù)免疫5周,成模大鼠出現(xiàn)與人類PM非常相似的骨骼肌病變,且成模率高、死亡率低,造模效果理想。2.用異種骨骼肌勻漿誘導免疫的方法建立的自身免疫性炎性肌病Lewis大鼠模型,在出現(xiàn)與ⅡM患者相似的骨骼肌病變同時合并相當比例的肺間質(zhì)病變,且病理類型與ⅡM臨床十分相似,顯示本模型可為研究ⅡM合并肺間質(zhì)病變提供一種良好的實驗工具,此發(fā)現(xiàn)在國內(nèi)、外相關領域中尚屬首次。3.自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型出現(xiàn)肺間質(zhì)病變與多種炎癥和致纖維化因子的作用有關,通過這些因子的進一步研究可能發(fā)現(xiàn)新的治療靶點。4.自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型的肺泡炎和間質(zhì)纖維化與局部過度激活的細胞凋亡過程和異�；钴S的免疫炎性反應有關,而TRAIL及其受體可能參與了局部異常病理改變。5.通過本研究證明糖皮質(zhì)激素可有效減少自身免疫性炎性肌病肺間質(zhì)病變Lewis大鼠模型肺組織中多種細胞因子的表達,對減輕肺間質(zhì)纖維化有顯著療效,特別是在早期應用可使用較小劑量達到理想治療效果。
[Abstract]:Objective : To improve the model of autoimmune inflammatory myopathic rats .
2 . To investigate the occurrence of pulmonary interstitial lesions in the Lewis rat model of autoimmune inflammatory disease , and to establish a stable model of Lewis rat model of pulmonary interstitial lesion of autoimmune inflammatory muscular disease ;
3 . To explore the pathogenesis of pulmonary interstitial lesions in Lewis rats with autoimmune inflammatory disease ;
4 . To investigate the expression and effect of TRAIL and its receptors on the lung interstitial lesions in Lewis rat model of autoimmune inflammatory disease : 5 . To explore the effect and mechanism of glucocorticoid on pulmonary interstitial lesions in Lewis rat model of autoimmune inflammatory disease .


Methods : Sprague - Dawley rats , Westar rats and Lewis rats were grouped according to the factors such as strain , sex , immune dose and so on .
2 . The Lewis rat model of autoimmune inflammatory myropathy was established by a multi - point subcutaneous injection immunization of rabbit skeletal muscle homogenate , and the pathological examination and immunohistochemistry of bilateral lung tissues were taken to observe the lung lesions .
3 . The expression level of TGF - 尾 , ctgf , ICAM - 1 , VCAM - 1 and CD163 in lung tissue of Lewis rat model was detected by immunohistochemical staining and computer image analysis method .
4 . The expression of TRAIL and its receptor in lung tissue of Lewis rat model was detected by immunohistochemical staining and computer image analysis method , and its correlation with pathological changes in lung tissues was analyzed .
5 . To compare the effects of different doses and different treatment origins on the lung fibrosis score of Lewis rat model and the expression level of TGF - 尾 , ctgf , ICAM , VCAM , and so on in local lung tissues .


The results showed that the success rate of Lewis rat model was the highest in three rat strains , and the experimental tolerance was the best . At the same immunization dose , the rate of adult rat model was higher than that in male group . At the dose of 1 Omg / kg to 12 mg / kg , the model rate and pathological score of model were obviously increased , but the mortality of rats was increased . At 4 锝，

本文編號：1864707