锌指蛋白A20突变体转基因小鼠脓毒症肺损伤的研究  

Pneumonic injuries of Zinc finger protein A20 mutant transgenic mice with experimental sepsis

作　　者：吴丽娟[1] 陈潇[1] 冯建男[2] 但刚[1] 龚杨彬[1] 曾平[1] 

机构地区：[1]成都军区总医院检验科,成都610083 [2]军事医学科学院基础医学研究所第四研究室,北京100850

出　　处：《第三军医大学学报》2010年第5期435-438,共4页Journal of Third Military Medical University

基　　金：全军医学科研"十一五"计划面上项目(06MA189)~~

摘　　要：目的了解锌指蛋白A20突变体(A20△)转基因小鼠脓毒症肺损伤的特点,探讨A20△对小鼠脓毒症肺损伤的影响。方法以A20△转基因脓毒症小鼠为动物模型(A20△转基因组),采用流式细胞仪测定肺组织单核巨噬细胞浸润,ELISA测定肺组织A20△、TNF-α、IL-1β水平,结合肺体指数和病理检查对炎症及其损伤程度进行鉴定。实验同时设LPS对照组(LPS组)和生理盐水对照组(对照组)。结果给予LPS后6、12、24h,A20△转基因组小鼠肺体指数较LPS组明显下降(P<0.01,P<0.05);A20△转基因组小鼠肺组织浸润单核/巨噬细胞数较LPS组明显下降(P<0.01)。给予LPS后3、6h,A20△转基因组小鼠肺组织炎症细胞因子TNF-α水平较LPS组明显下降(P<0.01);IL-1β水平也较LPS组明显下降(P<0.05,P<0.01);给予LPS后12、24h,TNF-α和IL-1β水平差异无统计学意义(P>0.05)。病理学观察表明A20△转基因组小鼠肺组织充血程度、肺泡积液及肺间质增厚程度均较LPS组轻。结论A20△转基因小鼠在一定程度上具有抵御LPS诱导性肺损伤的能力,其机制可能是通过抑制单核细胞向肺组织浸润和降低肺内炎症细胞因子水平实现的。Objective To investigate the pneumonic injury features of zinc finger protein A20 mutant （ A20△ ） transgenic mice with experimental sepsis, and approach the regulation roles of A20△ and underlying mechanism. Methods Totally 72 Kunming mice were randomly divided into 3 groups, control, LPS group and A20△ transgene group. The mice of the control group received an injection of 1.6 ml normal saline through tail vein, those from the LPS group underwent an injection of 1.6 ml normal saline and then 2.5 mg/kg LPS in 8 h later, while those in A20△ transgene group were injected with 10ug/1.6 ml plasmid pCAGGS-A20 A followed by an injection of 2.5 mg/kg LPS in 8 h after the first injection. The mice were killed in 3, 6, 12 and 24 h after LPS injection and their lungs were reseeted. The quantity of mononuclear macrophage infiltrating in the lung was detected by flow cytometry. The levels of A20 mutant, TNF-α and IL-1β were analyzed by ELISA. The lung-body index and pathologic examination were recruited to accredit lung injury. Results In A20△ transgene group, the lung-body indexes were decreased obviously than that of LPS Group in 6, 12 and 24 h after LPS injection （P 〈 0.05, 0.01 ）. The amount of mononuelear macrophage infiltrating in the lung were decreased obviously than that of LPS group （P 〈0.01 ）. In 3 and 6 h after injection LPS, the levels of TNF-α in pneumonic tissue were decreased obviously than that of LPS group （ P 〈 0.01 ）. The levels of IL-1β in pneumonic tissue were also decreased obviously than that of LPS group （P 〈 0.05, 0.01 ）. After injection of LPS for 12 and 24 h, there were no difference in the levels of TNF-α and IL-1β in A20△ transgene group and LPS group （ P 〉 0.05 ）. The results of pathologic examination indicated that the pneumonic injury of A20△ transgene group was milder than LPS Group. Conclusion A20△ transgenic mice withstands the pneumonie inflammatory damages in LPS-induced sepsis at some extent, and the mechanism may be through suppre

关 键 词：脓毒症 肺 创伤和损伤 锌指蛋白 小鼠 转基因 

分 类 号：R631[医药卫生—外科学] R322.35[医药卫生—临床医学]