地榆皂苷Ⅱ对耐甲氧西林金黄色葡萄球菌的抗菌活性及作用机制研究  

Study on antibacterial activity and mechanism of ziyuglycoside Ⅱ against methicillin-resistant Staphylococcus aureus

作　　者：张子桦 吴金婵 余咏诗 张淼菲 张凯嘉 曾芝璇 卓燕萍 杨烨健 陈鑫 ZHANG Zihua;WU Jinchan;YU Yongshi;ZHANG Miaofei;ZHANG Kaijia;ZENG Zhixuan;ZHUO Yanping;YANG Yejian;CHEN Xin(School of Medicine,Foshan University,Foshan 528225,China;School of Food Science and Engineering,Foshan University,Foshan 528225,China;Foshan Hospital of Traditional Chinese Medicine,Foshan 528225,China)

机构地区：[1]佛山科学技术学院医学院,广东佛山528225 [2]佛山科学技术学院食品科学与工程学院,广东佛山528225 [3]佛山市中医院,广东佛山528225

出　　处：《药物评价研究》2024年第1期103-108,共6页Drug Evaluation Research

基　　金：广东省基础与应用基础研究基金项目(2019A1515110092)。

摘　　要：目的 考察地榆皂苷Ⅱ对耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA)的体外抑菌活性以及抑菌机制。方法 肉汤稀释法检测地榆皂苷Ⅱ对16株MRSA的体外抑菌活性;比浊法和菌落计数绘制地榆皂苷Ⅱ对MRSA的生长曲线、杀菌曲线;通过DNA外泄实验,分析地榆皂苷Ⅱ对MRSA细胞壁膜通透性的影响;通过DAPI染色后的荧光强度分析,考察地榆皂苷Ⅱ对MRSA DNA和RNA的影响。结果 地榆皂苷Ⅱ对16株MRSA的最小抑菌浓度(MIC)范围为128~256μg·mL^(-1),最小杀菌浓度(MBC)范围为2~8mg·mL^(-1)。生长曲线结果显示,地榆皂苷Ⅱ质量浓度为1×MIC(256μg·mL^(-1))及以上时,生长抑制作用明显;杀菌曲线结果显示,地榆皂苷Ⅱ2×MIC、4×MIC作用1 h后对MRSA的抑制作用明显,8×MIC、16×MIC分别作用1、2 h后对MRSA起到了完全抑制的作用。2×MIC的地榆皂苷Ⅱ作用于MRSA后,胞外核酸大分子含量显著高于对照组,6 h后核酸外泄量较对照组高26.23%(P<0.05)。地榆皂苷Ⅱ作用于MRSA菌体后,MRSA DNA和RNA水平明显低于对照组(P<0.01、0.001)。结论 地榆皂苷Ⅱ对MRSA有良好的抑菌活性,可以通过抑制细菌核酸合成进而抑制MRSA的生长。Objective To investigate the in vitro antibacterial activity and mechanism of traditional Chinese medicine monomer ziyuglycosideⅡagainst methicillin-resistant Staphylococcus aureus(MRSA).Method The broth dilution method was used to detect the in vitro antibacterial activity of ziyuglycosideⅡagainst 16 MRSA strains.Draw the growth curve and bactericidal curve of ziyuglycosideⅡon MRSA using turbidimetry and colony counting.Analyze the effect of ziyuglycosideⅡon the permeability of MRSA cell wall membrane through DNA leakage experiments.By analyzing the fluorescence intensity after DAPI staining,the effect of ziyuglycosideⅡon MRSA DNA and RNA was investigated.Result The minimum inhibitory concentration range of ziyuglycosideⅡon 16 MRSA strains was 128—256μg·mL^(−1),with a minimum bactericidal concentration range of 2—8 mg·mL^(−1).The growth curve results showed that when the mass concentration of ziyuglycosideⅡwas 1×MIC(256μg·mL^(−1))and above,the growth inhibition effect was significant.The sterilization curve results show that after one hour of 2×MIC and 4×MIC action,the inhibitory effect on MRSA was significant.ZiyuglycosideⅡof 8×MIC,16×MIC completely inhibited MRSA after 1 and 2 hours of action,respectively.After MIC induced action on MRSA,the content of extracellular nucleic acid macromolecules was significantly higher than that of the control group.After six hours,the amount of nucleic acid leakage was 26.23%higher than that of the control group(P<0.05).After the action of ziyuglycosideⅡon MRSA cells,the levels of MRSA DNA and RNA were significantly lower than those of the control group(P<0.01,0.001).Conclusion ZiyuglycosideⅡis an antibacterial drug with good antibacterial activity against MRSA,which can inhibit the growth of MRSA by inhibiting bacterial nucleic acid synthesis.

关 键 词：耐甲氧西林金黄色葡萄球菌 地榆皂苷Ⅱ 抗菌 细胞壁膜通透性 核酸合成 

分 类 号：R285.5[医药卫生—中药学]