实验性败血症小鼠模型的建立及评价  

Establishment and Evaluation of Experimental Sepsis Mouse Model

作　　者：王立燕[1] 徐若男[1] 韩根成[1] 王仁喜[1] 陈国江 肖鹤[1] 侯春梅[1] 沈倍奋[1] 黎燕[1] 

机构地区：[1]军事医学科学院基础医学研究所分子免疫研究室,北京100850

出　　处：《中国实验血液学杂志》2010年第3期766-770,共5页Journal of Experimental Hematology

基　　金：国家973项目(编号2007CB512406;2009CB522408);国家自然基金项目(编号30801029)

摘　　要：白血病、恶性淋巴瘤、再生障碍性贫血等造血系统恶性疾病,在应用化疗或大量的免疫抑制剂之后常常合并严重感染,使得近年来败血症的发生逐年增多,败血症早期诊断困难,死亡率高。本研究建立成熟的实验性败血症小鼠模型,为探讨败血症中的致病机制提供体实验基础。以经典的盲肠结扎穿孔(CLP)方法建立实验性败血症小鼠模型,用ELISA法检测补体C5a、IL-6、TNF-α、IFN-γ的水平,流式细胞术检测胸腺和肠系膜淋巴细胞的凋亡,HE染色方法观察胸腺和脾的病理损伤。实验结果显示,70%-80%左右的动物于术后72小时内死亡,在限定的观察时间内只有20%左右动物生存,CLP手术组的动物体重也有明显降低,补体C5a、IL-6、TNF-α、IFN-γ等与败血症相关的炎症介质在CLP手术后均表现为显著的上调,CLP术后20小时小鼠胸腺及肠系膜的CD4+淋巴细胞均出现了明显凋亡,胸腺及脾组织也发生了明显的病理损伤,研究结果显示了动物模型的可靠性和可行性。结论:本研究成功建立了败血症小鼠模型(CLP),为进一步探讨恶性血液病并发败血症的发病机制及干预策略提供了必要条件。After treating with chemotherapy or immunosuppressant, malignant diseases of hematopoietic system such as leukemia, malignant lymphoma and aplastic anemia usually induced severe infection such as sepsis. Sepsis which is hard to be diagnosed causes high death rate. This study was purposed to establish an experimental sepsis mouse model so as to provide a basis for pathogenesis and intervention study. A classic ceacal ligation and puncture （CLP） was used to establish experimental sepsis model. ELISA was used to detect levels of C5a, IL-6, TNFα, and IFN-γ. Flow Cytometry was applied to measure apoptosis of lymphocytes in thymus and mesentery. The pathologic changes of thymus and spleen were confirmed by HE staining. The results showed that almost 70% -80% mice died at 72 hours after CLP. Only approximate 20% animal suvived during finite time, mice in CLP group had significant weight lose. Meanwhile large release of different inflammatory mediators which are related with sepsis （CSa, IL-6, TNF-α, and IFN-γ） was observed after CLP. Apoptosis of lymphocytes in thymus and mesentery lymphonodes was enhanced markedly after CLP. Signifi- cantly pathologic injury was also observed in thymus and spleen. It is concluded that a mouse model of experimental sep- sis was successtully established by ceacal ligation and puncture which can well mimic the clinical symptom of sepsis. The experimental sepsis mouse model provides an excellent tool for exploring the pathogenesis and intervention ways for sepsis accompanied with complicated malignant hematological diseases in vivo.

关 键 词：败血症 盲肠结扎 盲肠穿孔 模型评价 

分 类 号：R631.3[医药卫生—外科学] R733[医药卫生—临床医学]