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(xué)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(jī)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(dá)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(jī)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(diǎn)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(jìn)行組織病理學(xué)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(biāo)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(diǎn)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(xué)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(jī)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(dá)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(jī)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(dá),并分析其與肺組織病理改變之間的相關(guān)性；5.比較不同劑量、不同治療起點(diǎn)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(dá)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(shí)驗(yàn)?zāi)褪苄宰詈?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(xì)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(yīng)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(xué)染色發(fā)現(xiàn)增厚肺泡間隔內(nèi)可見大量CD8+T淋巴細(xì)胞,支氣管及血管周圍可見大量CD20+B淋巴細(xì)胞；3.免疫組化染色結(jié)合計算機(jī)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(dá)量明顯高于對照組,特別是血管和支氣管周圍的陽性反應(yīng)最為明顯。同時ICAM和VCAM的表達(dá)量明顯高于對照組。CD163陽性細(xì)胞數(shù)明顯高于對照組。4.免疫組化染色結(jié)合計算機(jī)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(dá)明顯升高、DCR1表達(dá)明顯降低。同時肺間質(zhì)病變中浸潤淋巴細(xì)胞上部分高表達(dá)DCR1。相關(guān)性分析發(fā)現(xiàn)肺泡炎性反應(yīng)程度與肺組織中CD8+T淋巴細(xì)胞的浸潤有關(guān),肺纖維化程度與肺組織中TRAIL的表達(dá)升高有關(guān)；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(yù)則效果更優(yōu),其機(jī)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(guān)因子TRAIL和DR4的表達(dá)有關(guān)。 [結(jié)論]1.在應(yīng)用異種動物骨骼肌勻漿多點(diǎn)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(shí)驗(yàn)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(dǎo)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(dāng)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(shí)驗(yàn)工具,此發(fā)現(xiàn)在國內(nèi)、外相關(guān)領(lǐng)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(guān),通過這些因子的進(jìn)一步研究可能發(fā)現(xiàn)新的治療靶點(diǎn)。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(xì)胞凋亡過程和異�；钴S的免疫炎性反應(yīng)有關(guān),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(xì)胞因子的表達(dá),對減輕肺間質(zhì)纖維化有顯著療效,特別是在早期應(yīng)用可使用較小劑量達(dá)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