Zhang Caixia, Song Jie, Xu Yang, Wu Juan, Sun Weibin, Li Kuanyu
Department of Cell Biology and Genetics, Medical School of Nanjing University, Jiangsu Key Laboratory of Molecular Medicine, Nanjing 210093, China.
Zhonghua Kou Qiang Yi Xue Za Zhi. 2014 Oct;49(10):614-8.
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.
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.
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.
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.
通过检测促红细胞生成素产生肝细胞受体相互作用蛋白B2(Ephrin B2)及其受体对单核细胞THP - 1与人脐静脉内皮细胞(HUVEC)之间黏附的影响,探讨牙龈卟啉单胞菌(Pg)感染介导增强两者黏附的机制。
将PgATCC33277在厌氧罐中培养,THP - 1细胞分别以感染复数(MOI)为1:100用不同浓度的Pg感染8小时和24小时,检测THP - 1细胞中Ephrin B2受体的表达。去除游离的Pg后,将THP - 1细胞与HUVEC(有无EphrinB2过表达)共培养24小时,再额外培养23小时后检测HUVEC细胞中Ephrin B2的表达。
Pg感染后THP - 1细胞与HUVEC的黏附增强。Pg感染24小时后,THP - 1表达的Ephrin B2受体包括EphB3(5.169±0.152,P = 0.005)、EphB4(11.040±1.195,P = 0.001)和EphA4(4.976±0.122,P = 0.001)以及HUVEC表达的Ephrin B2(8.938±0.962,P = 0.008)的mRNA水平均显著升高。HUVEC中Ephrin B2的过表达促进了THP - 1与HUVEC的黏附。
Ephrin B2及其受体参与了Pg感染介导的THP - 1与HUVEC细胞黏附增强,提示Ephrin B2参与动脉粥样硬化的发展。