The KLF15 Regulate Downstream Genes With Deep Venous Thrombosis In The Venous Endothelial Cells Of Mice

DVT in the clinical departments,especially in orthopedics,have a high morbidity,but the diagnosis and treatment is difficult at early time,and have a high mortality rate when it is complicated by pulmonary embolism.Our research group has establish rat DVT model,we found that venous endothelial cell activation,coagulation/anticoagulation system imbalance and inflammatory reaction had close relationship with DVT.In this research we study the relationship and regulatory relationship between KLF15,and downstream genes with DVT in animal model.In animal model research we discuss the expression and functions of KLF15 and the downstream genes to investigate the association between the down stream genes and KLF15 with DVT.Objective:Established inferior vena cava stenosis DVT C57 mice model,after 24 hours we harvested inferior vena cava venous tissue,observed the DVT formation,detected the KLF15 expression and use the RNA-seq technology to the whole genome analysis of the differences genes then analysis the relationship between these genes with DVT.Method:1.60 C57 mice were randomly divided into blank control group(n=20)?DVT group(n=20),and siKLF15-DVT group(20),we inject the 0.9%NS 0.2ml to the control group and DVT group,KLF15 siRNA 10Onmol per mice through the tail vein,after 24 hours built models by stenosis inferior vena cava harvested inferior vena cava venous tissue 24 hours after stenosis,observed the DVT formation,we detected KLF15 expression of venous tissue by real-time PCR.2.Using RNA_seq technology analysis the difference genes between blank control group and DVT group,DVT group and siKLF group within the scope of the whole genome,discuss the relationship between these genes with DVT.Result:1.DVT group thrombosis rate 73%,motality rate 15%,thrombus wet weight 6.65±2.37mg,siKLF15 group thrombosis rate 88%,motality rate 10%,thrombus wet weight 15.85±2.64mg,real-time PCR detected mice venous tissue KLF15 mRNA expression in DVT groups and siKLF15 group.KLF15 expression were lower in siKLF15 group compare with DVT group(P<0.05).2.there are 15978 difference genes to be find between blank control group and DVT group(P<0.05),and have 701 significant difference genes(I log2(a fold change)|>1)at further screening.Among of those genes 497 genes are up-regulation expression and 204 genes down-regulation expression.However,there are 218 difference genes between siKLF15 group and DVT group(P<0.05),and have 88 significant difference genes(| log2(a fold change)|>1)at further screening.Among of those genes 20 genes are up-regulation expression and 68 genes down-regulation expression.Selected 17 gene in two group the in common.Strip2?Cd51.A2m.Ankrdl may regulat the IL-I?1?IL-6?TNF-a?TGF-?1 to activate the NF-? B pathways thus influence DVT.Conclusion:1.KLF15 can inhibit the formation of deep vein thrombosis;2.KLF15 may regulate downstream genes such as Strip2?CD51?A2m?Ankrd1 to influence of deep vein thrombosis;...