Milward M R, Chapple I L C, Wright H J, Millard J L, Matthews J B, Cooper P R
Periodontology, School of Dentistry, University of Birmingham, Birmingham B4 6NN, UK.
Clin Exp Immunol. 2007 May;148(2):307-24. doi: 10.1111/j.1365-2249.2007.03342.x. Epub 2007 Mar 9.
To investigate the molecular effects of the periodontopathogens Fusobacterium nucleatum (FN) and Porphyromonas gingivalis (PG) on the oral epithelium, the H400 oral epithelial cell line was cultured in the presence of non-viable bacteria. Following confirmation of the presence of transcripts for the bacterial pattern recognition receptors in H400 cells, Toll-like receptors -2, -4 and -9, and components of the NF-kappaB signalling pathway, immunocytochemical analyses were performed showing that NF-kappaB was activated within 1 h of exposure to both periodontopathogens. A significantly greater number of NF-kappaB nuclear translocations were apparent following H400 cell exposure to FN as compared with PG. Gene expression analyses indicated that transcripts known to be regulated by the NF-kappaB pathway, including cytokines/chemokines TNF-alpha, IL-1beta, IL-8, MCP-1/CCL2 and GM-CSF, were up-regulated following 4 and 24 h of exposure to both periodontopathogens. In addition, H400 periodontopathogen exposure resulted in differential regulation of transcripts for several cytokeratin gene family members. Consistent with the immunocytochemical data, microarray results indicated that FN induced a greater number of gene expression changes than PG following 24 h of exposure, 609 and 409 genes, respectively. Ninety-one genes were commonly differentially expressed by both periodontopathogens and represented biological processes commonly associated with periodontitis. Gene expression analyses by reserve transcriptase-polymerase chain reaction (RT-PCR) of molecules identified from the microarray data sets, including Heme oxygenase-1, lysyl oxidase, SOD2, CCL20 and calprotectin components, confirmed their differential expression profiles induced by the two periodontopathogens. FN and PG have clearly different molecular effects on oral epithelial cells, potentially highlighting the importance of the composition of the plaque biofilm in periodontitis pathogenesis.
为了研究牙周病原体具核梭杆菌（FN）和牙龈卟啉单胞菌（PG）对口腔上皮细胞的分子影响，在存在非存活细菌的情况下培养H400口腔上皮细胞系。在确认H400细胞中存在细菌模式识别受体、Toll样受体-2、-4和-9以及NF-κB信号通路的成分的转录本后，进行免疫细胞化学分析，结果显示在暴露于这两种牙周病原体后1小时内NF-κB被激活。与PG相比，H400细胞暴露于FN后，NF-κB核转位的数量明显更多。基因表达分析表明，已知受NF-κB途径调节的转录本，包括细胞因子/趋化因子肿瘤坏死因子-α、白细胞介素-1β、白细胞介素-8、单核细胞趋化蛋白-1/CCL2和粒细胞-巨噬细胞集落刺激因子，在暴露于这两种牙周病原体4小时和24小时后上调。此外，H400细胞暴露于牙周病原体导致几种细胞角蛋白基因家族成员的转录本受到不同调节。与免疫细胞化学数据一致，微阵列结果表明，暴露24小时后，FN诱导的基因表达变化比PG更多，分别为609个和409个基因。两种牙周病原体共同差异表达的91个基因代表了通常与牙周炎相关的生物学过程。通过逆转录聚合酶链反应（RT-PCR）对从微阵列数据集中鉴定出的分子进行基因表达分析，包括血红素加氧酶-1、赖氨酰氧化酶、超氧化物歧化酶2、CCL20和钙卫蛋白成分，证实了它们由两种牙周病原体诱导的差异表达谱。FN和PG对口腔上皮细胞具有明显不同的分子影响，这可能突出了菌斑生物膜组成在牙周炎发病机制中的重要性。
