CAJ | 학술논문

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.
목적 검측간배단백B2 (erythropoietin producing hepatomocellular receptor interacting protein B2,Ephrin B2)급기수체재단핵세포THP-1점부인제정맥내피세포(human umbilical vein endothelial cell,HUVEC)중적작용,게시아간계람단포균(Porphyromonas gingivalis,Pg)감염증강THP-1점부HUVEC적분자궤제.방법 응용염양관배양Pg,이감염복수1∶100감염인단핵세포주THP-1,감염8화24 h후,수집부분양품분별용우검측THP-1세포Ephrin B2급기수체표체수평적변화;기여양품여전염공재체혹과표체Ephrin B2적HUVEC세포공배양,용실시형광정량화단백질인적법검측HUVEC세포Ephrin B2적표체수평,병관찰Ephrin B2대THP-1여HUVEC점부적영향.결과 Pg감염24 h후,THP-1세포Ephrin B2수체EphB3、EphB4화EphA4적mRNA표체수평분별제고지5.169±0.152、11.040± 1.195화4.976±0.122,여미감염대조조(결과수거위1)상비차이균유통계학의의(P＜0.01);여경Pg감염적THP-1공배양후,HUVEC세포Ephrin B2적표체제고지8.938±0.962,여미감염대조조상비차이균유통계학의의(P=0.008);불경과감염,HUVEC직접과표체Ephrin B2후,THP-1대HUVEC점부성현저증가.결론 감염Pg적THP-1점부HUVEC적수량증다여Ephrin B2급기수체적상호작용유관,제시동맥혈관내피세포특이성표체적Ephrin B2분자삼여동맥죽양경화적발생.
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.