Study On The Role Of Endothelial Cells Derived Factors HMGB-1 And KLF-2 In The Early Stage Of DVT Formation

The causes of deep vein thrombosis are varied, so the previous diagnosis methods cannot be adopted in the disease determination of early stage. The difficulties of pre-diagnosis severely affect the promptness of treatment. If the patients cannot be treated timely and effectively in the early stage, they may suffer from further pathological injuries in limbs, and even the symptoms of dysfunction in limb veins. For the severe patients, the thrombosis may fall off to lung tissues, causing pulmonary artery embolism and even mortal risk. Therefore, it has been the discussion focus for both domestic and foreign researchers to study the relative marks of preventing and pre-diagnosing DVT.In recent years, among several causes of DVT, the endothelial cell injuries gradually become the heating point of DVT causes. The inflammatory factors released by injured endothelial cells may stimulate white cells and activate platelets, thus causing and accelerating the formation of thrombosis. KLF-2、HMGB-1 are closely related with endothelial cells which are the key role in DVT formation; KLF-2、 HMGB-1 inhibits thrombosis through protecting the stability of endothelial cells and negatively regulating the functions of inflammatory factors. This study focuses on KLF-2、HMGB-1, establishes DVT animal models, observe the expression changes on of above mentioned marks in DVT formation process through molecular biology testing methods, and initially discuss the inner relationship of the two factors and their role in deep vein thrombosis formation.Objective:1. Establish C57 DVT model of mice with inferior vena cava stenosis method, observe thrombosis formation status with pathological sections, and get inferior vena cava and thrombosis tissues of mice;2. Detect the expression changes of KLF-2 and HMGB-1 on the transcription facet in inferior vena cava and thrombosis tissues by real-time PCR; initially discuss its relationship with DVT and its roles.In order to provide basic experimental evidence for further analyzing about the relationship KLF-2 and PAI-1 with DVT.3. Using ELISA method to examine the expression of t-PA in plasma, to discuss the relationship between DVT and t-PA. In order to provide basic experimental evidence for further analyzing about the relationship between t-PA and DVT.Methods1. Establish DVT model of mice, get sections and detect:120 C57 mice are divided randomly into control group (n=40), sham operation group (n=40) and DVT group (n=40). Within 24 hours upon modeling,1.5cm inferior vena cava tissues are taken from renal vein in control group and sham operation group, while inferior vena cava tissues and its contents are taken within the scope of thrombosisin DVT group.Then datas were recorded such as inferior vena cava diameter.2. Make pathological sections of inferior vena cava tissues, observe thrombosis formation after HE staining; detect mRNA expression level of KLF-2 and HMGB-1 in vein tissues by real-time PCR and analyze their relationship with DVT formation.3. Using ELISA method to examine the expression of t-PA in plasma of each group. To discuss the role it may play in DVT formation.Results1. HE staining of vein tissues in DVT group:complete thrombosis can be seen in DVT group, inflammatory cells infiltration within vascular wall is obvious; no thrombosis forms in control group and sham operation group.2. Detect mRNA expression level of KLF-2 in vein tissues by real-time PCR: expression level of DVT group increases compared with control group and sham operation group, with statistic significance (p<0.05); the comparison between control group and sham operation group is without statistic significance (p>0.05).The mRNA expression level of HMGB-1:expression level of DVT group decreases compared with control group and sham operation group, with statistic significance (p<0.05); the comparison between control group and sham operation group is without statistic significance (p>0.05).3. Using ELISA method to examine the expression of t-PA:expression level of DVT group increases compared with control group and sham operation group, with statistic significance (p<0.05); the comparison between control group and sham operation group is without statistic significance (p>0.05).Conclusion1. In the mice model of DVT,the change of KLF-2、HMGB-1、t-PA expression indicate that, they are associated with DVT.In the Early Stage of DVT Formation, they may affect the formation of thrombosis.2. KLF-2、HMGB-lmay affect the expression of t-PA To break the dynamic balance of fibrinolytic-fibrinolytic resistance system, and play a role in inhibiting thrombosis.