Gene expression of human coronary artery endothelial cells in response to Porphyromonas endodontalis invasion

Hee-Joung Kong; Kyoung-Kyu Choi; Sang-Hyuk Park; Jin-Yong Lee; Gi-Woon Choi

doi:10.5395/JKACD.2009.34.6.537

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Original Article Gene expression of human coronary artery endothelial cells in response to Porphyromonas endodontalis invasion: Hee-Joung Kong¹, Kyoung-Kyu Choi¹, Sang-Hyuk Park¹, Jin-Yong Lee², Gi-Woon Choi¹; Journal of Korean Academy of Conservative Dentistry 2009;34(6):537-550.
DOI: https://doi.org/10.5395/JKACD.2009.34.6.537
Published online: November 30, 2009

¹Department of Conservative Dentistry, Division of Dentistry, Graduate of Kyung Hee University, Korea.

²Institute of Oral Biology, Division of Dentistry, Graduate of Kyung Hee University, Korea.

Corresponding Author: Gi-Woon Choi. Professor of Division of Dentistry, Graduate school of KyungHee University, 1, Hoegi Dong, Dongdaemun Gu, Seoul, Korea, 130-702. Tel: 82-2-958-9336, gwchoi@khu.ac.kr

• Received: September 8, 2009 • Revised: September 28, 2009 • Accepted: October 28, 2009

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Abstract
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Abstract

During the last two decades, there has been an increasing interest in the impact of oral health on atherosclerosis and subsequent cardiovascular disease (CVD). To date, some periodontal pathogens including Porphyromonas gingivalis (P. gingivalis) have been reported to be relevant to CVD. Porphyromonas endodontalis (P. endodontalis), which shares approximately 87% sequence homology with P. gingivalis, is mostly found within infected root canals. However, recent studies reveal that this pathogen also resides in the dental plaque or periodontal pocket in patients with periodontitis. It has been shown that P. endodontalis invades human coronary artery endothelial cells (HCAEC) and coronary artery smooth muscle cells (CASMC). To evaluate whether P. endodontalis can participate in the progression of atherosclerosis and CVD, we examined the changes in transcriptional gene expression profiles of HCAEC responding to invasion by P. endodontalis in this study.

The following results were obtained.
1. Porphyromonas endodontalis was invasive of HCAEC.
2. According to the microarray analysis, there were 625 genes upregulated more than two-folds, while there were 154 genes downregulated by half.
3. Upregulated genes were relevant to inflammatory cytokines, apoptosis, coagulation and immune response. Enhanced expression of MMP-1 was also noticeable.
4. The transcription profiles of the 10 selected genes examined by real-time PCR agreed well with those observed in the microarray analysis.
Thus, these results show that P. endodontalis presents the potential to trigger and augment atherosclerosis leading to CVD.
Keywords: Porphyromonas endodontalis; Atherosclerosis; Cardiovascular disease (CVD); Invasion; Human coronary artery endothelial cells (HCAEC); Gene expression

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Table 1

Primers and product sizes for RT-PCR analysis^a

^aThe full names of the genes and their UniGene IDs are given in Table 3.

Table 2

Invasion of human coronary artery endothelial cells by P. endodontalis strains

^aDefined as the actual number of viable cells of P. endodontalis inoculum as determined by plating method. Bacterial cell suspension was adjusted to optical density of 0.1 at 600nm and 100µl of the suspension was added to each well (1ml) of a 24-well plate.

^bDefined as the number and percentage of P. endodontalis cells protected from metronidazole killing, which represents invading bacterial cells, after the infection period. Values are means (± standard deviation) of triplicate.

Table 3

Upregulated genes relevant to CVD including atherosclerosis

Table 4

Comparison of gene expression measured by microarray and real-time PCR

Table 5

Pathway analysis of the significantly upregulated genes

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Gene expression of human coronary artery endothelial cells in response to Porphyromonas endodontalis invasion

J Korean Acad Conserv Dent. 2009;34(6):537-550. Published online November 30, 2009

DOI: https://doi.org/10.5395/JKACD.2009.34.6.537

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Gene expression of human coronary artery endothelial cells in response to Porphyromonas endodontalis invasion

Table 1

Primers and product sizes for RT-PCR analysis^a

^aThe full names of the genes and their UniGene IDs are given in Table 3.

Table 2

Invasion of human coronary artery endothelial cells by P. endodontalis strains

Table 3

Upregulated genes relevant to CVD including atherosclerosis

Table 4

Comparison of gene expression measured by microarray and real-time PCR

Table 5

Pathway analysis of the significantly upregulated genes

Table 1 Primers and product sizes for RT-PCR analysisa

^aThe full names of the genes and their UniGene IDs are given in Table 3.

Table 2 Invasion of human coronary artery endothelial cells by P. endodontalis strains

^aDefined as the actual number of viable cells of P. endodontalis inoculum as determined by plating method. Bacterial cell suspension was adjusted to optical density of 0.1 at 600nm and 100µl of the suspension was added to each well (1ml) of a 24-well plate.

^bDefined as the number and percentage of P. endodontalis cells protected from metronidazole killing, which represents invading bacterial cells, after the infection period. Values are means (± standard deviation) of triplicate.

Table 3 Upregulated genes relevant to CVD including atherosclerosis

Table 4 Comparison of gene expression measured by microarray and real-time PCR