大黄酸干预木糖葡萄球菌生物被膜及其作用机制研究  

Intervention and mechanism research of rhein on Staphylococcus xylosus biofilms

作　　者：吕婷婷 张忠斌 贺爽 马金鑫 丁文雅 Lü Ting-ting;ZHANG Zhong-bin;HE Shuang;MA Jin-xin;DING Wen-ya

机构地区：[1]广西中医药大学中药制剂共性技术研发重点实验室,广西南宁530200 [2]东北农业大学动物医学院,黑龙江哈尔滨150030

出　　处：《饲料研究》2022年第1期72-76,共5页Feed Research

基　　金：国家自然科学基金项目(项目编号:82060806);广西中医药大学引进博士科研启动基金项目(项目编号:2019BS006);广西中医药大学大学生创新创业训练项目(项目编号:202010600170) 中药学广西一流学科项目(项目编号:桂教科研[2018] 12号)。

摘　　要：试验通过标准微量稀释法测定大黄酸的亚抑菌浓度(MIC),利用结晶紫染色法考察大黄酸对木糖葡萄球菌生物被膜形成的干预作用;以木糖葡萄球菌谷氨酰胺酶缺失株和咪唑甘油磷酸酯脱水酶缺失株为研究对象,测定不同亚抑菌浓度药物大黄酸对各菌株生物被膜形成能力的影响,考察药物在生物被膜不同形成时期对细菌生物被膜形成能力的影响,探索大黄酸干预木糖葡萄球菌生物被膜形成的作用机制。结果显示,大黄酸对木糖葡萄球菌的MIC为64 mg/L;亚抑菌浓度的大黄酸具有抑制木糖葡萄球菌生物被膜形成的能力,且随着大黄酸浓度增加,其干预作用越明显,抑制效果越好;以木糖葡萄球菌标准菌株作为参照,谷氨酰胺酶缺失株、咪唑甘油磷酸酯脱水酶缺失株形成生物被膜的能力极显著下降(P<0.01);在各亚抑菌浓度大黄酸中谷氨酰胺酶缺失株抑制率均显著降低,咪唑甘油磷酸酯脱水酶缺失株在低浓度大黄酸中抑制率也显著降低;药物大黄酸对木糖葡萄球菌生物被膜的干预主要在6 h与12 h发挥显著作用,谷氨酰胺酶对生物被膜的抑制时间也在6 h和12 h。研究表明,亚抑菌浓度大黄酸可以显著抑制木糖葡萄球菌生物被膜形成,通过谷氨酰胺酶、咪唑甘油磷酸酯脱水酶为靶点发挥干预作用;且大黄酸对生物被膜的干预时期在生物被膜形成的早期阶段,可能是通过抑制细菌黏附发挥干预生物被膜形成作用。The minimum inhibitory concentration(MIC)of rhein was determined by standard microdilution method,and crystal violet staining method was used to investigate the intervention of rhein on the biofilm formation of S.xylosus.The S.xylosus of glutaminase deletion strains and the imidazole glycerol phosphate dehydratase deletion strains were used as the research object to measure the effect of different subinhibitory concentrations of rhein on the biofilm formation ability of each strain and to investigate the effects of drugs on the biofilm formation ability of bacteria at different biofilm formation stages.The experimental data were analyzed to explore the mechanism of rhein on biofilm formation of S.xylosus.The MIC of rhein on S.xylosus was 64 mg/L.The subinhibitory concentration of rhein could inhibit the biofilm formation of S.xylosus,and the inhibitory effect was better with the increase of rhein concentration.With S.xylosus as reference,the ability of forming biofilm of glutaminase deletion strains and imidazole glycerol phosphate dehydratase deletion strains decreased extremely(P<0.01),the inhibition rate of glutaminase deletion strains in each subinhibitory concentration of rhein was significantly decreased,and the inhibition rate of imidazole glycerol phosphate dehydratase deletion strains was significantly decreased in low concentration of rhein.The intervention of rhein on the biofilm of S.xylosus was mainly at 6 h and 12 h,and the inhibition time of glutaminase was also at 6 h and12 h.The experiment indicates that the subinhibitory concentration of rhein can significantly inhibit the biofilm formation of S.xylosus.Rhein inhibits the formation of S.xylosus biofilm is the result of the intervention of biofilm by multiple targets,including glutaminase and imidazole glycerol phosphate dehydratase,which play an intervention role.The intervention period of rhein on biofilm formation is in the early stage of biofilm formation,which may play a role in biofilm formation by inhibiting bacterial adhesion.

关 键 词：奶牛乳腺炎 大黄酸 木糖葡萄球菌 生物被膜 干预生物被膜形成机制 

分 类 号：S853.7[农业科学—临床兽医学]