链球菌E24无细胞上清液抗浮游态和生物被膜态金黄色葡萄球菌活性的研究  

Inhibitory effect of cell-free supernatant of Streptococcus E24 on planktonic and biofilm state Staphylococcus aureus

作　　者：李娜 孙冶[1,2] 李新鸣 高毅敏[1,2] 王鹏宇 孙贺 LI Na;SUN Ye;LI Xinming;GAO Yimin;WANG Pengyu;SUN He(Institute of Basic Medicine,Shenyang Medical College,Shenyang,Liaoning 110034,China;不详)

机构地区：[1]沈阳医学院基础医学院,辽宁沈阳110034 [2]辽宁省环境污染与微生态重点实验室 [3]沈阳医学院附属第二医院检验科

出　　处：《中国微生态学杂志》2025年第2期155-165,共11页Chinese Journal of Microecology

基　　金：沈阳医学院研究生科技创新基金项目(Y20240501);辽宁省创新能力提升联合基金(2022-NLTS-14-05);辽宁省教育厅科学研究项目(SYYX202012)。

摘　　要：目的探究链球菌E24无细胞上清液对浮游态金黄色葡萄球菌生长和生长过程中毒力因子释放的影响,以及对生物被膜形成的抑制和清除作用,为防治金黄色葡萄球菌感染、研制抗细菌感染的微生态制剂提供新的思路。方法采用牛津杯双层琼脂法和96微孔板法测定链球菌E24无细胞上清液对浮游态金黄色葡萄球菌生长的影响。测定链球菌E24无细胞上清液对浮游态金黄色葡萄球菌生长过程中溶血素、凝固酶及肠毒素表达量的影响。体外构建金黄色葡萄球菌生物被膜,采用结晶紫和刚果红染色法检测链球菌E24无细胞上清液对金黄色葡萄球菌生物被膜形成与胞外多糖产生的抑制,及其对成熟生物被膜的清除作用。应用光镜和电镜观察生物被膜的形态,并采用RT-qPCR法检测金黄色葡萄球菌生物被膜相关基因(icaA、cidA和sarA)表达水平的变化。结果链球菌E24无细胞上清液对浮游态金黄色葡萄球菌生长有显著抑制作用,可以降低浮游态金黄色葡萄球菌生长过程中溶血素、凝固酶及肠毒素的释放。链球菌E24无细胞上清液可以降低金黄色葡萄球菌的生物被膜形成量和胞外多糖产量,降低生物被膜相关基因的表达水平。结论链球菌E24无细胞上清液能抑制浮游态金黄色葡萄球菌生长和毒力因子的表达,并对其生物被膜态有良好的抑制和清除作用。Objective To explore the effects of Streptococcus E24(hereinafter the E24)cell-free supernatant(CFS)on the growth and release of virulence factors during the growth process of planktonic Staphylococcus aureus(S.aureus),and its inhibitory and clearing effects on biofilm formation,providing new ideas for the prevention and treatment of S.aureus infection and the development of microecological agents for bacterial infections.Methods The effect of E24 CFS on the growth of planktonic S.aureus was determined using the Oxford Cup double-layer agar method and 96 well plate method.The effect of E24 CFS on the expression levels of hemolysin,coagulase and enterotoxin during the growth process of planktonic S.aureus was determined.The S.aureus biofilm was constructed in vitro,the inhibitory and destructive effects of E24 CFS on the formation of S.aureus biofilm and the production of extracellular polysaccharides were measured by using crystal violet semiquantitative method and Congo red staining method.Morphological alteration of the biofilms was observed under light and electron microscopy,and RT-qPCR was used to detect the changes in the expression levels of S.aureus biofilm related genes(icaA,cidA and sarA).Results The E24 CFS had a significant inhibitory effect on the growth of planktonic S.aureus,which reduced the release of hemolysin,coagulase and enterotoxins during the growth process of planktonic S.aureus.E24 CFS reduced the biofilm formation and extracellular polysaccharide production of S.aureus,and lowered the expression levels of biofilm related genes.Conclusion E24 CFS can inhibit the growth of planktonic S.aureus and the expression of virulence factors during the growth process,and has a good inhibitory and clearing effect on its biofilm state.

关 键 词：链球菌E24 金黄色葡萄球菌 浮游态菌 生物被膜 拮抗作用 

分 类 号：R378.1[医药卫生—病原生物学]