Analysis of the interaction between periodontopathic bacteria and junctional epithelial cells using in situ (RT-) PCR

我们研究了牙周病细菌与牙周接合上皮细胞之间的相互作用。在RT-PCR中，暴露于牙周病艾克莱氏菌腐蚀物后，上皮细胞中IL-8、IL-6、PGHS-2 mRNA水平显著升高。有趣的是，与e.s rodens上清孵育后，KB细胞IL-8、IL-6、PGHS -2 mRNA水平显著升高。这些结果表明，口腔上皮细胞对IL-6、IL-8和前列腺素E2的模拟并不一定需要直接接触。我们认为，E.腐蚀诱导上皮细胞分泌促炎细胞因子，作为宿主结缔组织免疫细胞和炎症细胞的早期信号系统。为了阐明中性粒细胞通过上皮细胞迁移的机制，我们研究了上皮细胞对E.锈菌附着反应中粘附分子的表达。在RT-PCR中，ICAM-1 mRNA…More水平在暴露于E. crodeens 1小时内显著升高，并在12小时内持续升高。此外，用n -乙酰- d -半乳糖胺（GalNAc）抑制腐蚀乳杆菌对KB细胞的附着可降低ICAM-1的表达，纯化的腐蚀乳杆菌粘附素（GalNAc特异性凝集素样物质，EcLS）可提高上皮细胞上ICAM-1的表达。这些结果表明，EcLS在牙龈上皮细胞ICAM-1表达中起关键作用。为了阐明锈蚀菌感染KB细胞后的信号转导，我们检测了锈蚀菌附着在上皮细胞上被磷酸化的成分。western blot检测到p42/44MAK激酶磷酸化，但未检测到EGF受体磷酸化。在组织免疫学研究中，在上皮细胞中检测到p42/44MAP激酶磷酸化，但在结缔组织中未检测到。此外，MAP激酶抑制剂阻断了ICAM-1和IL-8的表达。综上所述，E. corrosion和EcLS分泌的蛋白可以通过p42/44 MAP激酶介导与上皮细胞相互作用。少

We examined the interaction between periodontopathic bacteria and periodontal junctional epithelial cells. In RT-PCR, Significant IL-8, IL-6, PGHS-2 mRNA levels were induced from epithelial cells in response to exposure to periodontopahtic Eiklenella corrodens . Interestingly, After incubation with E. corrodens supernatant, KB cells showed marked increase in the levels of IL-8, IL-6, PGHS -2 mRNA. These results imply that the direct contact of E. corrodens with oral epithelial cells is not necessarily required for the sirumulation pf IL-6, IL-8, and Prostaglamdin E2. We suggest that E. corrodens induces the epithelial cells to secrete proinflammatory cyotkines which serve as an early signaling system to host immune and inflammatory cells in underlying conective tissue.To elucidate the mechanism of the transmigration neutrophils through the epithelium, we investigated the expression of adhesion molecules on epithelial cells in response to E. corrodens attachment. In RT-PCR, ICAM-1 mRNA … More levels were significantly increased within 1h in response to exposure to E. corrodeens and continued over the 12h period of study. Furthermore, inhibition of the attachment of E. corrodens to KB cells with N-acetyl-D-galactosamine (GalNAc) decreased the expression of ICAM-1 and purified adhesin of E. corrodens (GalNAc specific lectin like substance, EcLS) increased the ICAM-1 expression on epithelial cells. These findings show that EcLS plays the key role in ICAM-1 expression on gingival epithelial cells.To clarify the signal transduction in KB cells after infection of E. corrodens, we examined components that were phosphrylated by the attachment of E. corrodens to epithelial cells. In western blot analysis, phosphorylated p42/44MAK kinase was detected but phosphorylated EGF receptor was not detected. In histoimmunologycal study, phosphorylated p42/44MAP kinase were detected in epithelium but not connective tissues. Furthermore, inhibitors of MAP kinase blocked the expression of ICAM-1 and IL-8. In summary, secreted proteins from E. corrodens and EcLS can interact with epithelial cells mediating with p42/44 MAP kinase. Less