深静脉血栓形成模型大鼠股静脉内皮组织中核转录因子kappa B1和组织因子的表达(英文)

Expression of nuclear factor kappa B1 and tissue factor in femoral venous endothelial tissue of the rat deep vein thrombosis model

背景:目前调控静脉内皮细胞、血小板、炎性细胞之间相互作用,促进局部深静脉血栓微环境形成的机制尚不完全清楚,仍未发现早期诊断深静脉血栓的可靠方法。目的:研究核转录因子kappaB1和组织因子在大鼠深静脉血栓形成中的作用。方法:将67只SD大鼠随机分为对照组和模型组,对模型组采用股静脉钳夹联合下肢石膏制动构建大鼠深静脉血栓形成模型。于造模后2.5,25h,解剖股静脉观测血栓的发生情况,进一步将模型组分为:血栓形成前组(造模后2.5h)、血栓形成组和血栓不形成组(造模后25h)。取各组股静脉内皮组织,采用基因芯片筛查差异表达的基因,并进一步采用real-timePCR进行验证。结果与结论:基因芯片分析及real-timePCR结果均发现,造模后2.5h,血栓形成前组大鼠股静脉内皮组织中核转录因子kappaB1和组织因子表达明显高于对照组(P<0.05);造模后25h,血栓形成组大鼠股静脉内皮组织中核转录因子kappaB1和组织因子表达明显高于血栓形成前组、血栓不形成组和对照组(P<0.05)。提示局部静脉内皮组织中核转录因子kappaB1和组织因子表达水平上调可能在深静脉血栓形成中发挥了重要作用。

BACKGROUND：At present,the basic underlying molecular mechanism regulating the interactions among venous endothelial cells,platelets,leukocytes,and promoting local deep vein thrombosis microenvironment formation,still remains unclear,and there is no ideal method for early diagnosis of deep vein thrombosis.OBJECTIVE：To study the underlying role of nuclear factor kappa B1 and tissue factor in rats with deep vein thrombosis.METHODS：A total of 67 Sprague-Dawley rats were randomly divided into control group（n=10） and model group（n=57）.Deep vein thrombosis model was established by a clamping and sewing method in femoral vein combined with cast fixation.The incidence and serious degree of thrombus were observed by dissecting rat femoral vein in different time points（2.5 and 25 hours after modeling）.The model group was further divided into pre-thrombogenesis group（2.5 hours after modeling）,thrombogenesis group（25 hours after modeling） and non-thrombogenesis group（25 hours after modeling）.Then total RNA was extracted from the localized femoral venous endothelial tissue.The candidate genes,associated inflammation and thrombosis,were screened by a special gene chip.Then the gene expression of nuclear factor kappa B1 and tissue factor was further identified by real-time polymerase chain reaction.RESULTS AND CONCLUSION：Pre-thrombogenesis group had no thrombogenesis,while thrombogenesis group have 23 cases with thrombosis and non-thrombogenesis group have 22 cases without thrombosis.The results of gene chip hybridization analysis and real-time PCR found that the mRNA expression of nuclear factor kappa B1 and tissue factor in rat femoral vein endothelial tissue were significantly up-regulated at 2.5 hours after modeling（pre-thrombogenesis group was higher than control group）（P 0.05）,and continued up-regulating at 25 hours after modeling（thrombogenesis group was higher than the pre-thrombogenesis group,non-thrombogenesis group and control group）（P 0.05）.The results from present study indicate that up-regulating expressions of nuclear factor kappa B1 and tissue factor in local femoral venous endothelial tissue of rat deep vein thrombosis models may play a key role in initiating venous thrombosis.