Minami Masaaki, Takase Hiroshi, Nakamura Mineo, Makino Toshiaki
Department of Bacteriology, Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japann.
Core Laboratory, Graduate School of Medical Sciences, Nagoya City University, Nagoya, Japan.
Drug Discov Ther. 2019;13(6):335-342. doi: 10.5582/ddt.2019.01087.
Foodborne diseases have become a worldwide problem that threatens public health and welfare. Enteropathogenic Escherichia coli (EPEC) is one of major pathogens of moderate to severe diarrhea. The increased prevalence of EPEC strains that produce extended spectrum β-lactamase (ESBL) has deepened the problem. The fruit of Lonicera caerulea var. emphyllocalyx (LCE) has been used as a traditional food preservative and medicine in northern temperate zones such as Hokkaido Island, Japan. In this study, we investigated the antibacterial effect of LCE fruit extract (LCEE) against EPEC. The antibacterial activities of LCEE were examined by bacterial growth, time-kill curve, soft-agar motility, electron microscopy, and 96 well-microplate biofilm assays. We also investigated the bacterial mRNA expression of biofilm-associated genes (fliC, csgA, and fimA) by quantitative real-time PCR assays. LCEE was found to suppress the growth, time-kill curve, and spread of EPEC. It also reduced the biofilm formation in a dose-dependent manner. Morphological analysis using transmission and scanning electron microscopy revealed that LCEE diminished the function of flagella resulting in reduced motility and biofilm formation. The mRNA expression of all three biofilm associated genes was downregulated under LCEE treatment. Extracts of the fruit of LCE inhibit the motility and biofilm formation of EPEC as a result of the inhibition of flagella development and function. We propose LCEE as a therapeutic candidate for the effective therapy of EPEC-associated infectious diseases.
食源性疾病已成为一个威胁公众健康和福祉的全球性问题。肠致病性大肠杆菌(EPEC)是导致中度至重度腹泻的主要病原体之一。产超广谱β-内酰胺酶(ESBL)的EPEC菌株患病率增加,使问题更加严重。蓝靛果忍冬(LCE)的果实已在日本北海道岛等北温带地区用作传统食品防腐剂和药物。在本研究中,我们调查了LCE果实提取物(LCEE)对EPEC的抗菌作用。通过细菌生长、时间-杀菌曲线、软琼脂运动性、电子显微镜和96孔微孔板生物膜试验检测了LCEE的抗菌活性。我们还通过定量实时PCR试验研究了生物膜相关基因(fliC、csgA和fimA)的细菌mRNA表达。结果发现LCEE可抑制EPEC的生长、时间-杀菌曲线和扩散。它还以剂量依赖的方式减少生物膜形成。使用透射和扫描电子显微镜进行的形态学分析表明,LCEE削弱了鞭毛的功能,导致运动性和生物膜形成降低。在LCEE处理下,所有三个生物膜相关基因的mRNA表达均下调。LCE果实提取物通过抑制鞭毛发育和功能来抑制EPEC的运动性和生物膜形成。我们建议将LCEE作为治疗EPEC相关传染病的有效治疗候选药物。