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肾上腺素和去甲肾上腺素对变异链球菌 UA159 生物膜形成和毒力因子的影响。

Effect of adrenaline and noradrenaline on biofilm formation and virulence factors of Streptococcus mutans UA159.

机构信息

State University of Campinas, School of Dentistry of Piracicaba, Piracicaba, SP, Brazil.

State University of Campinas, School of Dentistry of Piracicaba, Piracicaba, SP, Brazil; Graduate Program in Biological Sciences, Federal University of Alfenas, Alfenas, MG, Brazil.

出版信息

Arch Oral Biol. 2021 May;125:105091. doi: 10.1016/j.archoralbio.2021.105091. Epub 2021 Feb 23.


DOI:10.1016/j.archoralbio.2021.105091
PMID:33652302
Abstract

OBJECTIVES: To evaluate in vitro the effects of adrenaline and noradrenaline on the biofilm formation on orthodontic brackets, acid production and expression of virulence genes of Streptococcus mutans UA159 (S. mutans). DESIGN: S. mutans UA159 biofilm was formed on orthodontic brackets under exposure to adrenaline (100 μM), noradrenaline (50 μM) or PBS solution (control group) in triptone-yeast extract with 1 % sucrose. After 24 h, biofilm formation was quantified through Colony Forming Units / mL (CFU/mL) and RNA was extracted to perform gene expression analysis through real-time reverse transcriptase-PCR (RT-qPCR). Evaluation of acid production was carried out on planktonic cultures for 6 h. One-way ANOVA followed by Tukey's test was carried to determine statistical difference. The level of significance was set at 5 %. RESULTS: Catecholamines stimulated biofilm formation of S. mutans in orthodontic brackets (p < 0,05) but did not interfere with acid production (pH reduction) or the expression of the tested genes related to biofilm formation (gtfB, gtfC, gbpA, gbpB, gbpC, gbpD and brpA), aciduric (relA) and acidogenic properties (ldh). CONCLUSIONS: The present study was the first to demonstrate that catecholamines can stimulate S. mutans UA159 biofilm formation. These findings can contribute to clarify the role of stress on bacterial metabolism and contribute to the understanding of a possible role on caries development, mainly in orthodontic patients.

摘要

目的:评估肾上腺素和去甲肾上腺素对正畸托槽生物膜形成、产酸和变形链球菌 UA159(S. mutans)毒力基因表达的体外影响。

设计:在含有 1%蔗糖的三肽酵母提取物中,将 S. mutans UA159 生物膜暴露于肾上腺素(100 μM)、去甲肾上腺素(50 μM)或 PBS 溶液(对照组)下形成于正畸托槽上。24 小时后,通过集落形成单位/毫升(CFU/mL)定量生物膜形成,并提取 RNA 通过实时逆转录聚合酶链反应(RT-qPCR)进行基因表达分析。对浮游培养物进行 6 小时的产酸评估。采用单因素方差分析(ANOVA)和 Tukey 检验确定统计学差异。显著性水平设定为 5%。

结果:儿茶酚胺刺激正畸托槽中 S. mutans 的生物膜形成(p<0.05),但不干扰产酸(pH 值降低)或与生物膜形成相关的测试基因的表达(gtfB、gtfC、gbpA、gbpB、gbpC、gbpD 和 brpA)、耐酸(relA)和产酸特性(ldh)。

结论:本研究首次证明儿茶酚胺可以刺激 S. mutans UA159 生物膜形成。这些发现有助于阐明应激对细菌代谢的作用,并有助于理解应激在龋齿发展中的可能作用,特别是在正畸患者中。

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