Effects of Porphyromonas gingivalis with different fimA genotypes on vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 production by human umbilical vein endothelial cells

Department of Periodontology, Affiliated Stomatological Hospital, Zunyi Medical College, China.
Zhonghua kou qiang yi xue za zhi = Zhonghua kouqiang yixue zazhi = Chinese journal of stomatology 06/2011; 46(6):332-7.
Source: PubMed


To investigate the effect of Porphyromonas gingivalis (Pg) with different fimA genotypes on vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) production by human umbilical vein endothelial cells (HUVEC).
In the present study, PgATCC33277 (type I fimA genotype), WCSP 115 (type II fimA genotype), W83 (type IV fimA genotype), and Escherichia coli-lipopolysaccharide (Ec-LPS) were designed as experimental group 1, 2, 3, and positive control group, respectively, to stimulate HUVEC, and the un-stimulated HUVEC were analyzed as negative control group. The three strains of Pg were cultured anaerobically in standard condition, and then the Pg cells and Ec-LPS were co-cultured with HUVEC for 2, 6, and 24 h, respectively. The amount of ICAM-1 and VCAM-1 produced by HUVEC was detected with flow cytometry (FCM). The expression of ICAM-1 and VCAM-1 by HUVEC were assayed with confocal laser scanning microscope (CLSM).
The expression of ICAM-1 on the surface of HUVEC were intensified after infected by Pg with I, II, and IV fimA genotypes (P < 0.05). The amounts of ICAM-1 were 60.27 ± 5.43, 80.81 ± 1.44, and 85.94 ± 2.56 for Pg with type I fimA genotype, 86.69 ± 8.81, 90.19 ± 0.00, and 96.18 ± 0.48 for Pg with type II fimA genotype, 59.66 ± 0.40, 85.79 ± 4.86, and 96.04 ± 2.07 for Pg with type IV fimA genotype at 2, 6 and 24 h after infection, respectively. The up-regulation effects caused by Pg with type II and IV fimA genotypes were stronger than those caused by Pg with type I fimA genotype at different time points except at 2 h (P < 0.05). Under the present experimental condition, infected by Pg with type I, II and IV fimA genotypes stimulated low expression of VCAM-1 by HUVEC, it showed no significant differences among all the groups (P > 0.05). Expression of ICAM-1 and VCAM-1 in Pg infected HUVEC were confirmed by CLSM. Infection of HUVEC with Pg resulted in more fluorescence staining of ICAM-1 and VCAM-1 compared with that in uninfected HUVEC cultures.
The virulence and pathogenicity of Pg is associated with its fimA genotypes, Pg with type II and IV fimA genes possess stronger ability to stimulate HUVEC to up-regulate the expression of cell adhesion molecules, which may lead to disorders in vascular endothelial function.

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    ABSTRACT: Background and Objective Porphyromonas gingivalis has been shown to actively invade endothelial cells and induce vascular cell adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) overexpression. Nucleotide-binding oligomerization domain 1 (NOD1) is an intracellular pattern recognition reporter, and its involvement in this process was unknown. This study focused on endothelial cells infected with P. gingivalis, the detection of NOD1 expression and the role that NOD1 plays in the upregulation of VCAM-1 and ICAM-1.Material and Methods The human umbilical vein endothelial cell line (ECV-304) was intruded by P. gingivalis W83, and cells without any treatment were the control group. Expression levels of NOD1, VCAM-1, ICAM-1, phosphorylated P65 between cells with and without treatment on both mRNA and protein levels were compared. Then we examined whether mesodiaminopimelic acid (NOD1 agonist) could increase VCAM-1 and ICAM-1 expression, meanwhile, NOD1 gene silence by RNA interference could reduce VCAM-1, ICAM-1 and phosphorylated P65 release. At last, we examined whether inhibition of NF-κB by Bay117082 could reduce VCAM-1 and ICAM- 1 expression. The mRNA levels were measured by real-time polymerase chain reaction, and protein levels by western blot or electrophoretic mobility shift assays (for phosphorylated P65).ResultsP. gingivalis invasion showed significant upregulation of NOD1, VCAM-1 and ICAM-1. NOD1 activation by meso-diaminopimelic acid increased VCAM-1 and ICAM-1 expression, and NOD1 gene silence reduced VCAM-1 and ICAM-1 release markedly. The NF-κB signaling pathway was activated by P. gingivalis, while NOD1 gene silence decreased the activation of NF-κB. Moreover, inhibition of NF-κB reduced VCAM-1 and ICAM-1 expression induced by P. gingivalis in endothelial cells.Conclusion The results revealed that P. gingivalis induced NOD1 overexpression in endothelial cells and that NOD1 played an important role in the process of VCAM-1 and ICAM-1 expression in endothelial cells infected with P. gingivalis through the NF-κB signaling pathway.
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