裸鼠肢体缺血后炎性细胞浸润及相关因子表达的动态变化  

Dynamic changes of inflammatory cells infiltration and related gene expression after limb ischemia in nude mice

作　　者：李大勇[1] 郑巧楠[1] 袁明殿[1] 陈文娜[2] 谷峰[3] 吕延伟 

机构地区：[1]辽宁中医药大学附属医院血管外科,辽宁省沈阳市110032 [2]辽宁中医药大学教学实验中心,辽宁省沈阳市110032 [3]辽宁中医药大学中医基础教研室,辽宁省沈阳市110032

出　　处：《中国组织工程研究与临床康复》2010年第41期7665-7670,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30600843):疏肝活血法促血管生成治疗肢体缺血性疾病的实验研究;"十一五"国家科技支撑计划项目(2008BAI53B012);辽宁省科技厅博士启动基金资助项目(20061030)~~

摘　　要：背景:有研究表明组织缺血后可发生代偿性血管新生,但缺血后局部炎症反应的变化规律及与血管新生的关系至今尚不明确。目的:观察裸鼠肢体缺血后组织中炎性细胞浸润及相关因子表达的动态变化。方法:用股动脉结扎法建立裸鼠肢体缺血模型,于缺血后3d,1,2,3,4周观察裸鼠肢体缺血状态的改变,应用苏木精-伊红染色、免疫组织化学染色分别观察缺血组织一般形态学、微血管数量和巨噬细胞浸润的变化,应用Westernblot和RT-PCR检测核因子κB、单核细胞趋化蛋白1和血管内皮生长因子表达的动态变化。结果与结论:裸鼠肢体缺血程度于造模后一两周最为严重,缺血后的肌肉纤维萎缩、变形,单核细胞趋化蛋白1和血管内皮生长因子表达最高(P<0.01)。微血管数量于缺血后2周时最多(P<0.01)。造模后3d～2周,缺血组织中有大量炎性细胞浸润,部分裸鼠的肢体出现缺血性坏疽,尤以缺血2周时最严重,核因子κB表达增强(P<0.01)。结果提示,核因子κB和单核细胞趋化蛋白1基因介导的炎症反应是肢体缺血发生后的代偿性变化,刺激了血管内皮生长因子的基因表达和短暂的血管新生过程,但由于微血管的生成数量有限,尚不足以代偿肢体缺血坏疽的发生。BACKGROUND： Research has shown that compensatory angiogenesis may occur after ischemia, but the relation between the changes of the local inflammatory response and angiogenesis after ischemia has rarely reported. OBJECTIVE： To discuss the dynamic changes of the inflammatory cells infiltration and related gene expression in muscle after limb ischemia. METHODS： Femoral artery ligation method was used to establish nude mice right hind limb ischemia model. The ischemic changes of limb ischemia were observed at 3 days, 1, 2, 3 and 4 weeks after limb ischemia. Hematoxylin-eosin staining, CD34 and CD68 immunohistochemical staining were used to observe ischemic morphological changes, the microvascular count （MVC） and macrophages infiltration in muscle tissue respectively. Western blot and reverse transcription polymerase chain reaction were used to detect changes of nuclear factor-kappa B （NF-κB）, monocyte chemotactic protein-1 （MCP-1） and vascular endothelial growth factor （VEGF） in ischemic muscle. RESULTS AND CONCLUSION： Limb ischemic state of nude mice was the most serious at 1-2 weeks after ischemia. After limb ischemia, muscle fibers shrink and deformation, and the expression of MCP-1 and VEGF were greatest （P0.01）. MVC was the highest at 2 weeks after limb ischemia （P0.01）. There were a large number of inflammatory cell infiltrations at 3 days-2 weeks, and parts of nude mice had ischemic gangrene, especially severe at 2 weeks, and the expression of NF-κB was enhanced （P0.01）. NF-κB and MCP-1 gene-mediated inflammatory response in limb ischemia was the compensatory changes, which stimulates VEGF gene expression and short-term process of angiogenesis. But the number of capillary formation is limited, thus, it was not enough to compensate the state of limb ischemic gangrene.

关 键 词：肢体缺血 动脉 炎性介质 基因表达 核因子ΚB 单核细胞趋化蛋白1 组织构建 

分 类 号：R318[医药卫生—生物医学工程]