KLF6通过上调TGF-β1诱导静脉内皮细胞endoglin,P-selectin表达,引发血小板粘附、聚集,促进TDVT形成的实验研究  

KLF6 upregulates endoglin,P-selectin expression and promotes thromgenesis by regulating TGF-β1 in vein endothelial cell

作　　者：李文[1] 胡继红[2] 李兴国[3] 李宏昆[3] 周如丹[3] 赵学凌[3] 王兵[3] 

机构地区：[1]云南省第一人民医院心内科,昆明650032 [2]昆明医学院第一附属医院放射科,昆明650032 [3]昆明医学院第一附属医院骨科,昆明650032

出　　处：《中国矫形外科杂志》2011年第16期1365-1368,共4页Orthopedic Journal of China

基　　金：国家自然基金资助项目(编号:30960389和81060151);云南省科技厅昆明医学院联合项目(编号:2009cd159)

摘　　要：[目的]研究创伤性深静脉血栓形成早期,大鼠静脉内皮细胞KLF6、TGF-β1表达上调,诱导Endog-lin、P-selectin表达,引发血小板粘附、活化、聚集,促进血栓形成的作用。[方法]将50只SD大鼠随机分为A组(对照组)、B组(血栓形成前组,创伤后2.5 h)、C组(血栓形成组,创伤后25 h)、D组(血栓不形成组,创伤后25 h),采用股静脉钳夹联合下肢石膏制动构建大鼠TDVT模型。不同时间点解剖股静脉观测血栓的发生率及严重程度。分离血管内皮,先采用Genechip Rat Genome 230 2.0基因芯片检测静脉内皮细胞中的基因表达变化,然后采用实时荧光定量聚合酶链式反应(real-time PCR)验证股静脉内皮中KLF6、Endoglin、TGF-β1、P-selectin表达;再对上述基因进行Pathway等生物信息学分析。[结果]C组大鼠(血栓形成)17例,D组大鼠(血栓未形成)13例,解剖发现C组内皮损伤较D组严重。基因芯片分析及real-time PCR结果均提示:创伤后2.5 h,KLF6,TGF-β1、Endoglin、P-selectin表达上调(Ratio值>2),B组高于A组,血栓形成时KLF6、TGF-β1、Endoglin、P-selectin表达显著上调(Ratio值>4),C组高于B、D组。Pathway分析提示:KLF6为TGF-β1和Endoglin的上游调控基因,TGF-β1为P-se-lectin的调控基因,P-selectin触发血小板粘附、活化、聚集及血栓形成。[结论]KLF6可通过上调TGF-β1诱导静脉内皮细胞endoglin、P-selectin表达,引发血小板粘附、聚集,促进大鼠深静脉血栓形成。[Objective]To study the effects of Krüppel-like Factors in increasing the expression of endoglin,P-selectin and promoting thrombosis by upregulating TGF-β1 in rats endothelial cells.[Method]The 50 SD rats were randomly divided into control group（group A,n = 10）,prethrombogenesis group（group B,2.5 hour after trauma）,thrombogenesis group（group C,25 hour after trauma） and non-thrombogenesis group（group D,25 hour after trauma）.TDVT（traumatic deep vein thrombosis） rat model were established by combination with clamping both femoral vein and fixing with cast.Rat femoral venous were dissected and observed the incidence and serious degree of thrombus in different time points.Then total RNA were extracted from the localized venous endothelial.After a gene chip-based screening,the gene expression of KLF6,endoglin,TGF-β1 and P-selectin was further identified by real-time PCR（real-time quantitative polymerase chain reaction）.[Result]There were 17 thrombosis in group C（thrombogensis group）,and 13 with non-thrombosis in group D（thrombogenesis group） in all model group.The injury degree of endothelial was more serious in group C than in group C by dissecting femoral vein.The results of gene chip hybridization analysis and real-time PCR confirmed that the mRNA expressions of KLF6,endoglin,TGF-β1 and P-selectin in group B were significantly increased（Ratio〉2） compared with group A in 2.5 hour after trauma（P〈0.05）.The mRNA expression of KLF6,endoglin,TGF-β1 and P-selectin in group C were significantly increased（Ratio〉4） compared with group B and D in 25 hour after trauma（P〈0.05）.Pathway analysis showed that KLF6 was the upstream regulatory genes of TGF-β1 and endoglin.TGF-β1 can regulate the expression of P-selectin.P-selectin can triggered platelet adhesion,activation,aggregation,and promote thrombosis.[Conclusion]KLF6 can upregulate Endoglin,P-selectin expression and promote thromgenesis by regulating TGF-β1 in vein endothelial cell.

关 键 词：KLF6 ENDOGLIN TGF-Β1 P-SELECTIN TDVT 基因芯片杂交技术 实时荧光定量聚合酶链式反应 

分 类 号：R543.6[医药卫生—心血管疾病]