Graziano T S, Closs P, Poppi T, Franco G C, Cortelli J R, Groppo F C, Cogo K
Department of Physiological Sciences, Area of Pharmacology, Anesthesiology and Therapeutics, Piracicaba Dental School, University of Campinas, Piracicaba, SP, Brazil.
J Periodontal Res. 2014 Oct;49(5):660-9. doi: 10.1111/jre.12148. Epub 2013 Nov 20.
Stress has been identified as an important risk factor in the development of many infectious diseases, including periodontitis. Porphyromonas gingivalis, a gram-negative oral anaerobic bacterium, is considered an important pathogen in chronic periodontitis. Microorganisms, including P. gingivalis, that participate in infectious diseases have been shown to respond to catecholamines released during stress processes by modifying their growth and virulence. Therefore, the purpose of this study was to evaluate the effects of adrenaline and noradrenaline on the growth, antimicrobial susceptibility and gene expression in P. gingivalis.
P. gingivalis was incubated in the presence of adrenaline and noradrenaline (100 μm) for different time-periods in rich (Tryptic soy broth supplemented with 0.2% yeast extract, 5 μg/mL of hemin and 1 μg/mL of menadione) and poor (serum-SAPI minimal medium and serum-SAPI minimal medium supplemented with 5 μg/mL of hemin and 1 μg/mL of menadione) media, and growth was evaluated based on absorbance at 660 nm. Bacterial susceptibility to metronidazole was examined after exposure to adrenaline and noradrenaline. The expression of genes involved in iron acquisition, stress oxidative protection and virulence were also evaluated using RT-quantitative PCR.
Catecholamines did not interfere with the growth of P. gingivalis, regardless of nutritional or hemin conditions. In addition, bacterial susceptibility to metronidazole was not modified by exposure to adrenaline or noradrenaline. However, the expression of genes related to iron acquisition (hmuR), oxidative stress (tpx, oxyR, dps, sodB and aphC) and pathogenesis (hem, hagA and ragA) were stimulated upon exposure to adrenaline and/or noradrenaline.
Adrenaline and noradrenaline can induce changes in gene expression related to oxidative stress and virulence factors in P. gingivalis. The present study is, in part, a step toward understanding the stress-pathogen interactions that may occur in periodontal disease.
压力已被确认为包括牙周炎在内的多种传染病发生发展的重要风险因素。牙龈卟啉单胞菌是一种革兰氏阴性口腔厌氧菌,被认为是慢性牙周炎的重要病原体。包括牙龈卟啉单胞菌在内的参与传染病的微生物已被证明可通过改变其生长和毒力来响应应激过程中释放的儿茶酚胺。因此,本研究的目的是评估肾上腺素和去甲肾上腺素对牙龈卟啉单胞菌生长、抗菌敏感性及基因表达的影响。
牙龈卟啉单胞菌在富含营养(添加0.2%酵母提取物、5μg/mL血红素和1μg/mL甲萘醌的胰蛋白胨大豆肉汤)和营养匮乏(血清-SAPI基础培养基以及添加5μg/mL血红素和1μg/mL甲萘醌的血清-SAPI基础培养基)的培养基中,于肾上腺素和去甲肾上腺素(100μm)存在的情况下孵育不同时间段,基于660nm处的吸光度评估生长情况。在暴露于肾上腺素和去甲肾上腺素后检测细菌对甲硝唑的敏感性。还使用逆转录定量聚合酶链反应评估参与铁摄取、应激氧化保护和毒力的基因表达。
无论营养或血红素条件如何,儿茶酚胺均不干扰牙龈卟啉单胞菌的生长。此外,暴露于肾上腺素或去甲肾上腺素不会改变细菌对甲硝唑的敏感性。然而,暴露于肾上腺素和/或去甲肾上腺素会刺激与铁摄取(hmuR)、氧化应激(tpx、oxyR、dps、sodB和aphC)及致病机制(hem、hagA和ragA)相关的基因表达。
肾上腺素和去甲肾上腺素可诱导牙龈卟啉单胞菌中与氧化应激和毒力因子相关的基因表达发生变化。本研究部分朝着理解牙周疾病中可能发生的应激-病原体相互作用迈出了一步。