Chou Hsin-Hua, Yumoto Hiromichi, Davey Michael, Takahashi Yusuke, Miyamoto Takanari, Gibson Frank C, Genco Caroline A
School of Dentistry, Taipei Medical University, Taipei, Taiwan.
Infect Immun. 2005 Sep;73(9):5367-78. doi: 10.1128/IAI.73.9.5367-5378.2005.
Epidemiological and pathological studies have suggested that infection with the oral pathogen Porphyromonas gingivalis can potentiate atherosclerosis and human coronary heart disease. Furthermore, infection with invasive, but not noninvasive P. gingivalis has been demonstrated to accelerate atherosclerosis in apolipoprotein E-deficient (ApoE(-/-)) mice and to accelerate local inflammatory responses in aortic tissue. In the present study, using high-density oligonucleotide microarrays, we have defined the gene expression profile of human aortic endothelial cells (HAEC) after infection with invasive and noninvasive P. gingivalis. After infection of HAEC with invasive P. gingivalis strain 381, we observed the upregulation of 68 genes. Genes coding for the cytokines Gro2 and Gro3; the adhesion molecules intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule (VCAM)-1, and ELAM-1 (E-selectin); the chemokine interleukin-8 (IL-8); and the proinflammatory molecules IL-6 and cyclooxygenase-2 were among the most highly upregulated genes in P. gingivalis 381-infected HAEC compared to uninfected HAEC control. Increased mRNA levels for signaling molecules, transcriptional regulators, and cell surface receptors were also observed. Of note, only 4 of these 68 genes were also upregulated in HAEC infected with the noninvasive P. gingivalis fimA mutant. Reverse transcription-PCR, enzyme-linked immunosorbent assay, and fluorescence-activated cell sorting analysis confirmed the expression of ICAM-1, VCAM-1, E-/P-selectins, IL-6, and IL-8 in HAEC infected with invasive P. gingivalis. We also demonstrated that increased expression of ICAM-1 and VCAM-1 in aortic tissue of ApoE(-/-) mice orally challenged with invasive P. gingivalis but not with the noninvasive P. gingivalis fimA mutant by immunohistochemical analysis. Taken together, these results demonstrate that P. gingivalis fimbria-mediated invasion upregulates inflammatory gene expression in HAEC and in aortic tissue and indicates that invasive P. gingivalis infection accelerates inflammatory responses directly in the aorta.
流行病学和病理学研究表明，感染口腔病原体牙龈卟啉单胞菌可加剧动脉粥样硬化和人类冠心病。此外，已证实感染具有侵袭性而非非侵袭性的牙龈卟啉单胞菌可加速载脂蛋白E缺陷（ApoE(-/-)）小鼠的动脉粥样硬化，并加速主动脉组织中的局部炎症反应。在本研究中，我们使用高密度寡核苷酸微阵列，确定了侵袭性和非侵袭性牙龈卟啉单胞菌感染后人主动脉内皮细胞（HAEC）的基因表达谱。用侵袭性牙龈卟啉单胞菌菌株381感染HAEC后，我们观察到68个基因上调。编码细胞因子Gro2和Gro3、黏附分子细胞间黏附分子1（ICAM-1）、血管细胞黏附分子（VCAM）-1和ELAM-1（E-选择素）、趋化因子白细胞介素-8（IL-8）以及促炎分子IL-6和环氧合酶-2的基因是牙龈卟啉单胞菌381感染的HAEC中与未感染的HAEC对照相比上调程度最高的基因。还观察到信号分子、转录调节因子和细胞表面受体的mRNA水平增加。值得注意的是，在感染非侵袭性牙龈卟啉单胞菌fimA突变体的HAEC中，这68个基因中只有4个也上调。逆转录-聚合酶链反应、酶联免疫吸附测定和荧光激活细胞分选分析证实了侵袭性牙龈卟啉单胞菌感染的HAEC中ICAM-1、VCAM-1、E-/P-选择素、IL-6和IL-8的表达。我们还通过免疫组织化学分析证明，用侵袭性牙龈卟啉单胞菌而非非侵袭性牙龈卟啉单胞菌fimA突变体经口攻击的ApoE(-/-)小鼠主动脉组织中ICAM-1和VCAM-1的表达增加。综上所述，这些结果表明牙龈卟啉单胞菌菌毛介导的侵袭上调了HAEC和主动脉组织中的炎症基因表达，并表明侵袭性牙龈卟啉单胞菌感染直接加速了主动脉中的炎症反应。
