IL-6mRNA Expression And Role In DVT Mouse And Human Blood

[Objective][1] Based on preliminary experiments on this subject DVT vein endothelial cells in rats differential gene screening results:IL-6gene differentially expressed genes in determining the expression of IL-6gene in the blood of DVT mice and human blood.[2] By pubmed, keeg and other biological information database, analysis of the relationship between IL-6key role in thrombosis endothelial cells, leukocytes, coagulation and fibrinolysis system, and explore them possible between DVT formation of specific links.[3] Combined with DVT mouse, human serum IL-6gene expression with specific bioinformatics analysis results, discussion whether IL-6formation associated with DVT, such as IL-6is associated with DVT diagnostic molecular markers as possible, in order to find the new diagnosis of deep vein thrombosis molecules provide the basis.[Materials and methods]1、104Kunming mice were randomly divided into control group (n=8, no further treatment), sham operation group (48, ligation of the inferior vena cava) and model group (48, dissect the inferior vena cava), set the purpose of the sham-operated groupexclusive traumatic Stress IL-6levels in the blood. The control group were randomly collected blood0.7-lml, sham group and model group in2h,4h,6h,8h,12h,24h time point each into8mice, blood samples were collected at each time point through the chambers of the heart (each mouse whole blood was collected0.7-lml); access to the model group, the inferior vena cava ligation at the distal end of the vascular tissue of about1.0cm and the same parts of the vascular tissue of control group. Observed by the naked eye and vascular tissue paraffin thrombosis extracted blood samples mRNA, RT-PCR, reverse transcription of the target gene mRNA to cDNA gene was amplified by PCR with GAPDH as an internal reference agar the sugar gel electrophoresis to detect changes in IL-6gene level to the Bio-Rad own the electrophoresis results software Quabtity one4.6analysis. Electrophoresis results of comparative statistical analysis, the study of different thrombus formation state level of IL-6gene changes in the occurrence and development of DVT.According to "the American College of chest physicians antithrombotic and thrombosis in clinical practice guidelines for the prevention and detection of deep venous thrombosis (2012Ninth Edition)" and "NICE" guide to prevent venous thromboembolism in the formulation of the diagnostic criteria of DVT:evaluation of comprehensive diagnosis=risk factors of DVT (basic risk factors and risk factors of main college) clinical manifestations and lower extremity deep venous color Doppler ultrasonography (Appendix2), a total of30patients with DVT, exclusion of other risk factors (oral contraceptives, venous vascular disease, obesity, diabetes, cancer, pregnancy, systemic inflammatory response syndrome, comprehensive factors of connective tissue disease)20patients included in the study, normal control group (20cases, healthy volunteers), post-traumatic without thrombosis group, trauma group (20cases, for Department of orthopedics trauma patients). Form three groups, gender and age, body mass index had no statistical significance (P>0.05). The three groups were in the morning fasting venous blood sampling, DVT group confirmed within7days from the date of acquisition, trauma group on the first postoperative day collection. PCR was used to detect the IL-6three gene expression changes in blood samples, compared with the animal experimental results, combined with the analysis of relation between activation of the molecule and the injury of endothelial cells, regulation of inflammation and coagulation system of internal and external sources such as Pubmed biological information retrieval platform. The molecular can be used as an early diagnosis of deep vein thrombosis marker analysis and evaluation.[2] To analyze the role of IL-6in thrombosis and the impossibility of IL-6to be a DVT diagnostic molecular combing bioinformatics methods and the experimental results[Result]1、Results of animal experiments:Model group2h no thrombosis,4h have incomplete thrombosis,6h have incomplete thrombosis,8h,12h,24h mice have complete thrombosis. control group without thrombosis; IL-6mRNA expression in each group (control group, sham operation group, model group), IL-6mRNA in each group (control group, sham operation group, model group) were expressed in sham group and the control group showed no significant difference (P>0.05); DVT model group at2h,4h,6h and the normal control group and the sham group, no statistically significant differential pressure (P>0.05),8h,12h,24h group compared with the sham group difference was statistically significant (P<0.05).Clinical trials of IL-6mRNA expression:IL-6mRNA expression in the three groups, the highest expression in the thrombosis group, the trauma the control group, followed by the healthy control group was the lowest, thrombosis group and the healthy control group and traumagroup difference was statistically significant (P<0.05).2、Bioinformatics analysis results:IL-6may be formed chemotaxis of monocytes/macrophages and endothelial cell adhesion and activation of platelets, endothelial cells, and other pathways involved in deep vein thrombosis.[Conclusion]IL-6and deep vein thrombosis are closely related, through chemokine monocyte-macrophage and venous endothelial cell adhesion, inhibiting fibrinolysis, endothelial cells, and other targets involved in the formation of DVT, IL-6has a DVT potential for molecular diagnostics.