缺氧诱导因子-1α和NADPH氧化酶在大鼠深静脉血栓形成中的作用  

Role of HIF-1α and NOX4 in rat deep venous thrombosis

作　　者：郑宏宇[1] 李兴国[1] 李宏昆[1] 赵学凌[1] 吴雪梅[1] 王兵[1] 

机构地区：[1]昆明医学院第一附属医院骨科,云南昆明650032

出　　处：《中国急救医学》2012年第1期35-39,共5页Chinese Journal of Critical Care Medicine

基　　金：国家自然科学基金资助项目（No.30960389,No.81060151,No.81160236）;云南省科技厅-昆明医学院联合项目（N0.2009CD159）;云南省重点新产品开发计划项目（No.2010BC010）;昆明医学院博士研究生创新基金项目（No.2011D07）

摘　　要：目的探讨大鼠深静脉血栓（DVT）模型股静脉内皮组织中缺氧诱导因子-1α（HIF-1α）和NADPH氧化酶（NOX4）的表达变化及其在血栓形成中的作用。方法将60只SD大鼠随机分为对照组（10只）和模型组（50只），对模型组采用股静脉钳夹联合双下肢石膏制动构建大鼠DVT模型。不同时间点（造模后2．5h和25h）解剖股静脉、观察血栓发生率，进而将模型组分为血栓形成前组（造模后2．5h）、血栓形成组（造模后25h）、血栓不形成组（造模后25h）。分离股静脉内皮组织，提取总RNA；采用Genechip Rat Genome2302．0基因芯片筛查差异表达的基因；采用实时荧光定量聚合酶链式反应（real—timePCR）验证这些基因的表达变化。结果基因芯片分析及real—timePCR结果均发现，大鼠股静脉内皮组织中HIF-1α和NOX4的表达水平，血栓形成组最高，血栓形成前组次之，均明显高于对照组和血栓不形成组（P〈0．05）。Pathway分析提示，缺氧条件下，HIF-1α可通过上调靶基因NOX4的表达，诱导活性氧（ROS）的产生，继而活化静脉内皮细胞，促进血栓形成。结论大鼠股静脉内皮组织中HIF-1α和NOX4表达上调，可能在创伤性DVT形成中发挥重要作用。Objective To study the role of hypoxia - inducible factor-1α （ HIF -1α） and NADPH oxidase （NOX4） in rat deep vein thrombosis （DVT） model Methods 60 SD rats were randomly divided into control group （ controlled blank, n = 10）, pre - thrombogenesis group （2.5 hours after the trauma）, thrombogenesis group （25 hours after the trauma） and non -thrombogenesis group （25 hours after trauma）. DVT rat models were established by clamping both femoral veins and cast fixing. The incidence and severity degrees of thrombus were observed by dissecting rat femoral veins at different time points. Then total RNA and proteins were extracted from the localized femoral venous endothelial tissues. After gene chip - based screen, the gene expressions of HIF -1α and NOX4 were further identified by real - time quantitative polymerase chain reaction（PCR）. Results The results of gene chip hybridization analysis and real - time PCR found that the mRNA expression of HIF -1α and NOX4 in rat femoral vein wall tissue significantly up - regulated at 2.5 hours after trauma （ pre - thrombogenesis group was higher than control group） （ P 〈 0. 05 ）, and continued up - regulating in thrombogenesis （thrombogenesis group was higher than pre -thrombogenesis group and non - thrombogenesis group） （P 〈 0.05 ）. Conclusion The results from present study indicate that increased expressions of HIF -1α and NOX4 in local femoral venous endothelial tissue may play a crucial role in DVT.

关 键 词：缺氧诱导因子-1Α NADPH氧化酶 静脉内皮组织 深静脉血栓 

分 类 号：R543.6[医药卫生—心血管疾病]