背景:目前,深静脉血栓的分子病因学机制及其形成的核心调控网络仍未完全阐明,对于深静脉血栓的早期诊断预测也无理想的方法。目的:研究组织蛋白酶L/G与创伤性深静脉血栓的预测。方法:采用蚊式钳夹闭50只SD大鼠双侧股静脉的3个不同部位3s随后予以模具制动制备大鼠创伤性深静脉血栓模型,根据股静脉血栓形成的不同阶段和生物学特征,将模型大鼠分为血栓形成前组、血栓形成组和无血栓形成组,另取10只正常大鼠作为对照组。在相应时间点取大鼠创伤静脉,提取总RNA,经过基因芯片技术筛选差异表达基因,并进一步应用real-time PCR进行验证。结果与结论:基因芯片杂交结果发现组织蛋白酶L/G基因在各组间差异表达明显,其中血栓形成组最高,无血栓形成组和血栓形成前组次之,均明显高于对照组(P〈0.05);real-time PCR分析结果与基因芯片杂交分析结果相一致。说明局部静脉血管壁中组织蛋白酶L/G表达水平升高与创伤性深静脉血栓形成有关,可作为深静脉血栓形成早期诊断、预测的候选分子标志。
BACKGROUND:At present,the basic molecular etiological mechanism and core regulatory network of deep vein thrombosis(DVT) remains uncertain,and there is not an ideal measure for early diagnosis of DVT.OBJECTIVE:To study the underlying impact of cathepsin L/G in DVT rat model.METHODS:DVT rat models(n = 50) were established by clamping both femoral vein in three different positions within 3 seconds with mosquito forceps and fixing with cast.According to different observation phases and biological situations of the femoral vein thrombosis,model rats were divided into thrombogenesis group,pre-thrombogenesis group and non-thrombogenesis group.An additional 10 normal rats served as control group.Femoral vein was obtained at corresponding time points to exact total RNA.After a gene chip-based screening,the data of gene expression were further dissected by real-time PCR.RESULTS AND CONCLUSUON:Gene chip hybridization analysis results demonstrated that differential expression of cathepsin L/G gene was significant among groups,and the expression was greatest in the thrombogenesis group,followed by pre-thrombogenesis and non-thrombogenesis groups,which was significantly greater than the control group(P 0.05).Real-time PCR analysis results were consistent with gene chip hybridization analysis results.These indicate that DVT is associated with an increase in expression of cathepsin L/G in local venous vascular wall,and they may be candidate molecular markers for early diagnosis of deep vein thrombosis.