luxS/AI-2密度感应对缓症链球菌生物膜致病力的影响
Biofilm formation and antimicrobial susceptibility of Streptococcus mitis in response to luxS/AI-2 signaling
作者：陆笑, 翁春辉, 王劲茗, 刘少娟, 刘芹, 林珊
Author：Lu Xiao, Weng Chunhui, Wang Jinming, Liu Shaojuan, Liu Qin, Lin Shan
收稿日期：2016-07-22 年卷（期）页码：2017,44(4):411-420
期刊名称：国际口腔医学杂志
Journal Name：International Journal of Stomatology
关键字：luxS基因,自体诱导物-2,密度感应,生物膜,抗菌药敏感性,
Key words：luxS gene,autoinducer-2,quorum sensing,biofilm,antimicrobial susceptibility,
基金项目：广州市科技计划应用基础研究专项项目（2011J4100036）
中文摘要
目的 了解luxS基因敲除,自体诱导物-2（AI-2）密度感应信号缺失对缓症链球菌生物膜致病力表型的影响。方法 常规细菌培养、药敏试验、形态学与生化反应鉴定,16S 核糖体DNA（rDNA）序列同源性分析,获得耐药性缓症链球菌临床分离株。同源重组法敲除luxS基因,构建突变株。哈氏弧菌BB170菌株生物发光实验检测AI-2信号;定量分析临床分离株与突变株生物膜形成量与抗菌药敏感性的差异;荧光黄染液、18909荧光增白染液、L7012死/活菌染液染色,激光共聚焦显微镜扫描,了解生物膜结构、胞外多糖、死/活菌分布差异;临床分离株上清液、4,5-二羟基-2,3-戊二酮（DPD）补偿生长实验确认突变株表型改变的原因。实验数据行Kolmogorov-Smirno非参数检验。结果 luxS基因突变株与临床分离株生物膜形成量存在显著性差异（P<0.001）;氨苄西林、环丙沙星、四环素干预条件下,突变株生物膜形成量显著降低。上清液、DPD补偿后,突变株生物膜形成能力和结构可以恢复。突变株生物膜结构变疏松,对氨苄西林、环丙沙星、四环素的敏感性增加。结论 luxS/AI-2密度感应信号缺失,导致耐药性缓症链球菌生物膜形成量下降,抗菌药敏感性增加,致病力表型发生改变。
英文摘要
ObjectiveThis paper aims to study the influence ofluxSgene mutation ofStreptococcus mitison biofilm formation and antimicrobial susceptibility.MethodsCulture in blood plate, Gram stain and susceptibility tests, and phy-siological and biochemical tests were first carried out. 16S ribosomal DNA(rDNA) sequence homology analysis of clinical isolate was examined subsequently. The autoinducer-2(AI-2) bioluminescence assay ofVibrio harveyiBB170 strain, multi-drug resistance, and ability of biofilm formation were examined inluxSmutant and clinical isolate. The structure, exopo-lysaccharides, and dead/live cells of biofilm were scanned by confocal laser scanning microscope. Compared with one another,luxSmutant growth recruited by isolate supernatant or 4,5-dihydroxy-2,3-pentanedione(DPD) was experimented.ResultsluxSgene deletion decreased biofilm formation(P<0 .001), loosened structure, and changed antimicrobial suscepti-bility(to ampicillin, ciprofloxacin, tetracycline). however, isolate supernatant or dpd can recover the structure and ability of biofilm formation inluxSmutant.ConclusionluxS/AI- 2 signaling can influence biofilm formation and antimicro-bial susceptibility ofS. mitiswith multidrug resistance.
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