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[不同fimA基因型牙龈卟啉单胞菌对人脐静脉内皮细胞血管细胞黏附分子-1和细胞间黏附分子-1产生的影响]

[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].

作者信息

Cai Shu-Yu, Lin Yu-Xiang, Xiao Li, He Quan-Min, Ge Song, Qian Min-Zhang

机构信息

Department of Periodontology, Affiliated Stomatological Hospital, Zunyi Medical College, China.

出版信息

Zhonghua Kou Qiang Yi Xue Za Zhi. 2011 Jun;46(6):332-7. doi: 10.3760/cma.j.issn.1002-0098.2011.06.001.

Abstract

OBJECTIVE

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).

METHODS

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).

RESULTS

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.

CONCLUSIONS

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.

摘要

目的

研究不同fimA基因型牙龈卟啉单胞菌(Pg)对人脐静脉内皮细胞(HUVEC)血管细胞黏附分子-1(VCAM-1)和细胞间黏附分子-1(ICAM-1)产生的影响。

方法

本研究中,将PgATCC33277(I型fimA基因型)、WCSP 115(II型fimA基因型)、W83(IV型fimA基因型)以及大肠杆菌脂多糖(Ec-LPS)分别设为实验组1、2、3和阳性对照组,用以刺激HUVEC,未受刺激的HUVEC作为阴性对照组进行分析。三株Pg在标准条件下厌氧培养,之后将Pg菌细胞和Ec-LPS分别与HUVEC共培养2、6和24小时。采用流式细胞术(FCM)检测HUVEC产生的ICAM-1和VCAM-1的量。用共聚焦激光扫描显微镜(CLSM)检测HUVEC中ICAM-1和VCAM-1的表达。

结果

I、II和IV型fimA基因型的Pg感染后,HUVEC表面ICAM-1的表达增强(P<0.05)。感染后2、6和24小时,I型fimA基因型Pg的ICAM-1量分别为60.27±5.43、80.81±1.44和85.94±2.56;II型fimA基因型Pg的ICAM-量分别为86.69±8.81、90.19±0.00和96.18±0.48;IV型fimA基因型Pg的ICAM-1量分别为59.66±0.40、85.79±4.86和96.04±2.07。除2小时外,不同时间点II型和IV型fimA基因型Pg引起的上调作用强于I型fimA基因型Pg(P<0.05)。在本实验条件下,I、II和IV型fimA基因型Pg感染刺激HUVEC低表达VCAM-1,各组间无显著差异(P>0.05)。CLSM证实了Pg感染的HUVEC中ICAM-1和VCAM-1的表达。与未感染的HUVEC培养物相比,Pg感染HUVEC导致ICAM-1和VCAM-1的荧光染色更多。

结论

Pg的毒力和致病性与其fimA基因型有关,II型和IV型fimA基因的Pg具有更强的刺激HUVEC上调细胞黏附分子表达的能力,这可能导致血管内皮功能紊乱。

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