自身免疫性炎性肌病Lewis大鼠模型肺间质病变及其机制的基础研究

发布时间：2018-05-09 05:08

  本文选题：自身免疫性炎性肌病 + 动物模型　； 参考：《北京协和医学院》2011年博士论文

【摘要】：[目的]1.完善自身免疫性炎性肌病大鼠模型的制作；2.探讨自身免疫性炎性肌病Lewis大鼠模型发生肺间质病变的情况,建立稳定的自身免疫性炎性肌病肺间质病变Lewis大鼠模型；3.探索自身免疫性炎性肌病Lewis大鼠模型肺间质病变的发生机制；4.探讨TRAIL及其受体在自身免疫性炎性肌病Lewis大鼠模型肺间质病变的表达和作用：5.探索糖皮质激素对自身免疫性炎性肌病Lewis大鼠模型肺间质病变的影响及其机制。 [方法]1.将SD大鼠、Westar大鼠和Lewis大鼠根据品系、性别、免疫物剂量等因素分组,通过兔骨骼肌匀浆等比例混合弗氏完全佐剂多点皮下注射免疫的方法建立自身免疫性炎性肌病大鼠模型,取股四头肌进行组织病理学检查,比较各组之间成模率、死亡率、肌肉病理评分等观察指标的差异；2.通过兔骨骼肌匀浆等比例混合弗氏完全佐剂多点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,取双侧肺行组织病理学检查和免疫组化检测,观察其肺部病变；3.通过免疫组化染色结合计算机图像分析方法检测自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺组织中TGF-β、CTGF、ICAM、VCAM及CD163等分子的表达水平,比较与对照组之间的差别；4.通过免疫组化染色结合计算机图像分析方法检测自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺组织中TRAIL及其受体的表达,并分析其与肺组织病理改变之间的相关性；5.比较不同剂量、不同治疗起点对自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺纤维化评分以及局部肺组织中TGF-β、CTGF、ICAM、VCAM等分子表达水平的影响。 [结果]1.三个大鼠品系中,Lewis大鼠模型成功率最高,且实验耐受性最好,相同免疫剂量下雌性组的成模率均高于雄性组,注射兔骨骼肌匀浆蛋白剂量从8mg/kg增加至1 Omg/kg时,成模率及模型病理评分明显升高,但从1 Omg/kg增加至12mg/kg时,成模率及模型病理评分无明显变化,而大鼠死亡率升高,免疫4～5周时可见骨骼肌纤维坏死、非坏死肌纤维及间质血管周围炎细胞浸润等典型病变；2.部分自身免疫性炎性肌病模型组Lewis大鼠肺组织出现不同程度肺间质病变表现,发生率为68.8%。典型的间质病变为间质性肺泡炎性反应和肺间质纤维化改变,最多见的病理类型为非特异性间质性肺炎(NSIP)和寻常型间质性肺炎(UIP),免疫组织化学染色发现增厚肺泡间隔内可见大量CD8+T淋巴细胞,支气管及血管周围可见大量CD20+B淋巴细胞；3.免疫组化染色结合计算机图像分析结果显示自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺组织中TGF-β和CTGF的表达量明显高于对照组,特别是血管和支气管周围的阳性反应最为明显。同时ICAM和VCAM的表达量明显高于对照组。CD163阳性细胞数明显高于对照组。4.免疫组化染色结合计算机图像分析方法显示自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺组织中TRAIL表达明显升高、DCR1表达明显降低。同时肺间质病变中浸润淋巴细胞上部分高表达DCR1。相关性分析发现肺泡炎性反应程度与肺组织中CD8+T淋巴细胞的浸润有关,肺纤维化程度与肺组织中TRAIL的表达升高有关；5.大剂量糖皮质激素可有效减轻自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺间质纤维化程度,小剂量、早干预则效果更优,其机制可能与减少病变局部TGF-β、CTGF和ICAM等炎症和促纤维化因子以及凋亡相关因子TRAIL和DR4的表达有关。 [结论]1.在应用异种动物骨骼肌匀浆多点皮下注方法建立自身免疫性炎性肌病大鼠模型时,选择雌性Lewis大鼠作为实验对象,给予兔骨骼肌匀浆1 Omg/kg连续免疫5周,成模大鼠出现与人类PM非常相似的骨骼肌病变,且成模率高、死亡率低,造模效果理想。2.用异种骨骼肌匀浆诱导免疫的方法建立的自身免疫性炎性肌病Lewis大鼠模型,在出现与ⅡM患者相似的骨骼肌病变同时合并相当比例的肺间质病变,且病理类型与ⅡM临床十分相似,显示本模型可为研究ⅡM合并肺间质病变提供一种良好的实验工具,此发现在国内、外相关领域中尚属首次。3.自身免疫性炎性肌病肺间质病变Lewis大鼠模型出现肺间质病变与多种炎症和致纤维化因子的作用有关,通过这些因子的进一步研究可能发现新的治疗靶点。4.自身免疫性炎性肌病肺间质病变Lewis大鼠模型的肺泡炎和间质纤维化与局部过度激活的细胞凋亡过程和异常活跃的免疫炎性反应有关,而TRAIL及其受体可能参与了局部异常病理改变。5.通过本研究证明糖皮质激素可有效减少自身免疫性炎性肌病肺间质病变Lewis大鼠模型肺组织中多种细胞因子的表达,对减轻肺间质纤维化有显著疗效,特别是在早期应用可使用较小剂量达到理想治疗效果。
[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，

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