Changes Of Gene HMOX-1 On Blood Prediction Diagnosis Deep Vein Thrombosis In Experiments

Objective To dynamically detect femoral vein gene expression changes in a rat model of traumatic deep vein thrombosis (TDVT) and to screen differential expression genes involved in this pathological process by gene chip detection method,Combined with Fold Change analysis, to screen differential expression genes from the genechips data of each group and to perform GO classification.Then to mainly analyze the expression changes of coagulation,anticoagulant,platelet associated genes HMOX-1 and so on.2 To dynamically detect HMOX-1 expression changes on blood in a rat model and hunman of traumatic deep vein thrombosis by RT-PCR method. In order to establish of new molecular markers to predict diagnosis of deep vein thrombosis in clinical provide experimental basis.[Materials and Methods]The first part:Summarized and analyzed the microarray data of traumatic deep vein thrombosis1 Combined with Fold Change analysis, to screen differential expression genes from the Affymetrix 230A gene chips detection data of each group and to perform GO classification.(Signal log2 ratio:comparison gene increases:Log2 Ratio≥1, gene discreases:Log2 Ratio≤-1)2Through an index word Coagulation,Anticoagulant,platelet in the NCBI website and CNKI database and screen significantly femoral vein gene expression changes of coagulation,anticoagulant,platelet associated genes and so on of through Fold Change analysis.(Signal log2 ratio comparison:gene increases:Log2 Ratio≥3, gene discreases:Log2 Ratio≤-3)3The selected the significant differentially expression changes of genes in the NCBI website GENE query its human and rat gene sequences, the two gene sequences into the BLAST than in a clear and homology of the human genome.The second part:HMOX-1 in rats predict the diagnosis of traumatic deep vein thrombosis experimental Study1 Directly clamping bilateral femoral veins combined with hip spica fixation was performed to establish TDVT rat models., According to build mode time and macro-scopic observation phases,100 SD rats were divided into 4 groups:the control (group A),initial period of thrombogenesis (group B),thrombogenesis at thrombotic crest-time (group C),nonthrombogenesis at thrombotic crest-time (group D).2 In the corresponding time, Every time,5ml peripheral blood were drawed of each rat, Obtain blood serum and blood cell specimen, total RNAs of them were extracted respe-ctively through TRIzol method.3RT-PCR were applied to detect expression levels of HMOX-1. At each phases, bilateral femoral veins were cut for HE staining and histological obervation in order to observe thrombosis phases.The third part:RT-PCR were applied to detect expression and Change of Gene HMOX-1 on blood in human of traumatic deep vein thrombosisAccording to the clinical observation, ultrasound, intravenous imaging. In the corresponding time, acquisition orthopaedic trauma and after surgery of deep vein thrombosis occurs and high-risk patients on blood Every time,5ml peripheral blood were drawed of each human, Obtain blood serum and blood cell specimen, total RNAs of them were extracted respectively through TRIzol method. RT-PCR were applied to detect expression levels of HMOX-1.[Results]1In this rat model, no thrombogenesis in A and D groups; to post-traumatic 24 h, The DVT rate of rat traumatic deep vein thrombosis model was 56.06%(37/66);The fem-oral vein macroscopic observation results of the groups were coherent to the histolo-gical observation results.2 In different phase point, expression changes level different of gene HMOX-1 in a rat modle of traumatic deep vein thrombosis,trend of raise express gradually; Both initial period of thrombogenesis (group B) and thrombogenesis at thrombotic crest-time (group C) compared with the control(groupA)were significant differences in up-regulated expression (BvsA:8.49,CvsA:14.76),while nonthrombogenesis at thrombotic crest-time (group D) compared with the control(groupA) were upregu-lated expression Slightly (DvsA:3.21);The RT-PCR and genechip detecting results of HMOX-1 presented a similar trends.3 The RT-PCR detecting results of HMOX-1:Gene of HMOX-1 no obvious differences in thrombosis and nothrombosis groups.[Conclusion]1 In different phase point, expression changes level different of gene HMOX-1 in a rat modle of traumatic deep vein thrombosis,trend of raise express gradually; The ratio HMOX-1 can indirectly reflect the different states of TDVT;2 HMOX-1 as predictive diagnostics deep vein thrombosis molecular markers possibility still need attempts and research in further.