肝配蛋白B2在牙龈卟啉单胞菌诱导单核细胞黏附人脐静脉内皮细胞中的作用  

Ephrin B2 is involved in Porphyromonas gingivalis infection-enhanced adhesion of THP-1 to human umbilical vein endothelial cells

作　　者：张彩霞[1] 宋洁[1] 徐杨[1] 吴娟[1] 孙卫斌[1] 李宽钰[1] 

机构地区：[1]南京大学医学院细胞生物学与遗传学教研室·江苏省医学分子技术重点实验室,210093

出　　处：《中华口腔医学杂志》2014年第10期614-618,共5页Chinese Journal of Stomatology

基　　金：南京大学医药生物技术国家重点实验室开放研究基金（KF-GN-201108）

摘　　要：目的 检测肝配蛋白B2 （erythropoietin producing hepatomocellular receptor interacting protein B2,Ephrin B2）及其受体在单核细胞THP-1黏附人脐静脉内皮细胞（human umbilical vein endothelial cell,HUVEC）中的作用,揭示牙龈卟啉单胞菌（Porphyromonas gingivalis,Pg）感染增强THP-1黏附HUVEC的分子机制.方法 应用厌氧罐培养Pg,以感染复数1∶100感染人单核细胞株THP-1,感染8和24 h后,收集部分样品分别用于检测THP-1细胞Ephrin B2及其受体表达水平的变化;其余样品与转染空载体或过表达Ephrin B2的HUVEC细胞共培养,用实时荧光定量和蛋白质印迹法检测HUVEC细胞Ephrin B2的表达水平,并观察Ephrin B2对THP-1与HUVEC黏附的影响.结果 Pg感染24 h后,THP-1细胞Ephrin B2受体EphB3、EphB4和EphA4的mRNA表达水平分别提高至5.169±0.152、11.040± 1.195和4.976±0.122,与未感染对照组（结果数据为1）相比差异均有统计学意义（P＜0.01）;与经Pg感染的THP-1共培养后,HUVEC细胞Ephrin B2的表达提高至8.938±0.962,与未感染对照组相比差异均有统计学意义（P=0.008）;不经过感染,HUVEC直接过表达Ephrin B2后,THP-1对HUVEC黏附性显著增加.结论 感染Pg的THP-1黏附HUVEC的数量增多与Ephrin B2及其受体的相互作用有关,提示动脉血管内皮细胞特异性表达的Ephrin B2分子参与动脉粥样硬化的发生.Objective To investigate the mechanisms of Porphyromonas gingivalis（Pg） infection-mediated enhancement of adhesion between monocytes THP-1 and human umbilical vein endothelial cells（HUVEC） by detecting the effect of erythropoietin producing hepatomocellular receptor interacting protein B2（Ephrin B2） and its receptors on the adhesion.Methods PgATCC33277 was cultured in an anaerobic jar,and THP-1 cells were infected with various concentrations of Pg at multiplicity of infection（MOI） of 1：100 for 8 and 24 h,respectively.The expression of Ephrin B2 receptor of THP-1 cells was detected.After removal of the free Pg,THP-1 cells were cocultured with HUVEC（overexpress of EphrinB2 or not） for 24 h to detect the expression of Ephrin B2 of HUVEC cells after additional cultivation for 23 h.Results The adhesion of THP-1 cells post infection by Pg to HUVEC was enhanced.The mRNA levels of Ephrin B2 receptors,including EphB3（5.169±0.152,P=0.005）,EphB4（11.040± 1.195,P=0.001）,and EphA4（4.976± 0.122,P=0.001） expressed by THP-1,and Ephrin B2（8.938±0.962,P=0.008） expressed by HUVEC were significantly elevated 24 h post infection of Pg.Over expression of Ephrin B2 in HUVEC promoted the adhesion of THP-1 to HUVEC.Conclusions Ephrin B2 and its receptors are involved in Pg infection mediated enhancement of the adhesion of THP-1 to HUVEC cells,suggesting that Ephrin B2 participates in the development of atherosclerosis.

关 键 词：紫单胞菌 龈 单核细胞 内皮细胞 促红细胞生成素产生的肝细胞受体-肝配蛋白B2 

分 类 号：R781.4[医药卫生—口腔医学]