The Research Of MicroRNA-181b Inhibit Deep Venous Thrombosis By Regulating P65 Expression In Vein Endotheliun

Deep venous thrombosis is a common disease of the cardiovascular system, its high incidence and complications pulmonary embolism is a serious threat to patients.However, the mechanism of deep vein thrombosis has not been studied clearly. The most representative of the theory was proposed by Virchow, which is the major factors of venous thrombosis, including blood stasis, vascular endothelial dysfunction and hypercoagulable state of blood. The common causes of vascular endothelial dysfunction include inflammation, oxidative stress, mechanical irritation and so on. In this study, the effects about miR-181b and NF-?B, VCAM-1, ICAM-1?E-selectin on deep vein thrombosis were studied. Through the animal modeling test and cell detection, the markers about deep vein thrombosis mechanisms is in-deep study.Objectives:1 SFP grade C57BL / 6 mice were used to model the stenosis, the changes of venous thrombosis after venous thrombosis model established were observed by comparing the percentage of thrombosis, thrombus diameter, thrombosis length and thrombosi wet weight in DVT group and in miR-181b overexpression or inhibition group.2 Measure the expression of VCAM-1, E-selectin and ICAM-1 in the venous endothelium by real-time PCR. Observe the activation of NF-?B signaling pathway.3 Investigate the effect of miR-181b on DVT formation and its mechanism.Methods:1 Use the SPF grade C57BL / 6 mice, average weigh is about 18-22g, the sex is not limited, The mice were randomly divided into 6 groups, each group include 20 mice.Normal group (Normal, n = 20), simple model group (DVT, n = 20), miRNA overexpression group (NS-m, n=20),miR-181b overexpression group (181 b-m, n =20), miRNA suppression group (NS-i, n = 20), miR-181b suppression group (181b-i,n = 20). Normal group and DVT group were tail injection with 0.2ml normal saline before 24h of the model establishment. NS-m group was tail injection with miRNA mimics 1nmol each mice before 24h of the model establishment, The NS-i group were tail injection with miRNA inhibitor 1 nmol each mice before 24h of the model establishment, the 181b-m group were tail injection with miR-181b-1 mimics 1 nmol each mice before 24h of the model establishment, the 181b-i group were tail injection of miR-181b-1 inhibitor 1 nmol each mice before 24h of the model establishment.MiRNA mimics, miRNA inhibitor, miR-181b-1 mimics, miR-181b-1 inhibitor were prepared with Lipofectamine 2000 as 0.2ml mixed liquid.2 Trate the mice with infrarenal vena cava (IVC) stenosis to establish the mice thrombosis model reference our group experience and classical literature. The mice were dissected at 24 hours after the model establishment, and observe the thrombosis each mouse. The expression of miR-181b and P65 in venous endothelium was identified by real-time PCR. The expression of VCAM-1, E-selectin and ICAM-1 were identified by real-time PCR.3 Analyze the experimental data by SPSS 22.0 statistical software. Discuss the effect of miR-181b on deep vein thrombosis and its possible mechanism.Results:1 The percentage of thrombosis, the thrombus diameter, thrombus length and thrombus wet weight of the vena cava thrombosis in mice with miR-181b overexpression group were lower than those in the DVT group. While when the expression of miR-181b was inhibited, the percentage. of thrombosis, the thrombus diameter, thrombus length, thrombus wet weight were significantly higher than the DVT group. The difference was statistically significant (P <0.05).2 The expression of VC AM-1, E-selectin and ICAM-1 was significantly lower than DVT group in the vena cava endothelium of mice after miR-181b overexpressioned,and the expression of VCAM-1, E-selectin , ICAM-1 were significantly higher than DVT group. The difference was statistically significant (P <0.05).3 The expression of P65 in mouse venous endothelium after miR-181b overexpressed was decreased compared with DVT group, the expression of P65 in venous endothelium was significantly increased compared with DVT group after miR-181b inhibited. The difference was statistically significant (P <0.05).Conclusions:1 miR-181b could inhibit the effect of venous thrombosis in mice DVT model.2 miR-181b could inhibit the expression of adhesion molecules like ICAM-1,VCAM-1, E-selectin in venous endothelium of DVT model.3 miR-181b could inhibit the expression of P65 in venous endothelium.4 miR-181b may inhibit the formation of DVT by inhibit the activation of NF-?B signaling pathway and inhibit the expression of adhesion molecules in venous endothelium.