薯蓣皂苷元对粪肠球菌体外抗菌性的研究

A study on diosgenin antibacterial activity against Enterococcus faecalis in vitro

目的:体外评价薯蓣皂苷元对粪肠球菌的抗菌效果。方法:体外培养粪肠球菌,将不同浓度薯蓣皂苷元与粪肠球菌共培养后,采用直接接触法和CCK-8法检测薯蓣皂苷元在0、1、5、10、50和100μmol/L时对粪肠球菌的抗菌性。采用菌落形成计数法(colony-forming units,CFU)检测薯蓣皂苷元在浓度为0、1、10、50和100μmol/L时活细菌数,并采用活死细菌染色法分析薯蓣皂苷元浓度为0,10和50μmol/L时粪肠球菌生物膜中活死细菌的比例。结果:细菌直接接触法和CCK-8法结果示薯蓣皂苷元在浓度为5、10、50和100μmol/L时,对粪肠球菌具有抗菌性,并呈浓度依赖性,50μmol/L时达到平台期。CFU计数法结果也显示薯蓣皂苷元在浓度为10、50和100μmol/L时对粪肠球菌具有显著的抗菌效果,50μmol/L时达到平台期。活死细菌染色法结果显示薯蓣皂苷元在浓度为10μmol/L和50μmol/L时,粪肠球菌生物膜活菌百分比显著下降,与阴性对照组相比差异有统计学意义。结论:薯蓣皂苷元对粪肠球菌具有良好的体外抗菌效果,将其作为根管消毒中的抗菌成分具有一定的临床应用前景。

Objective:To investigate the diosgenin antibacterial effects on Enterococcus faecalis(E.f)in vitro.Methods:E.f was cultured in vitro.After the co-culture of diosgenin and E.f,direct contact method and CCK-8 assay were used to detect diosgenin antibacterial activity at the different concentration of 0,1,5,10,50 and 100μmol/L.Bacterial colony forming unit(CFU)was used to detect the number of live bacteria at the different concentration of 0,1,10,50 and 100μmol/L.The ratio of live and dead bacteria in E.f biofilm was analyzed by staining with live and dead bacteria.Results:Compared with the control group,when the concentration reaches 5,10,50 and 100μmol/L(P<0.05),diosgenin had antibacterial properties and this phenomenon showed does-dependence.But there exists a plateau when the dosage beyond 50μmol/L.CFU counting method also showed the same trend and convinced that diosgenin had significant antibacterial effect against E.f when the concentration was 10,50 and 100μmol/L(P<0.05).The staining results of live and dead bacteria further confirmed that when the concentration of diosgenin was 10 and 50μmol/L,the percentage of viable bacteria in E.f biofilm decreased significantly compared with the control group(P<0.05).Conclusion:Diosgenin has a good antibacterial effect on E.f in vitro.It has a certain clinical application prospect as an antibacterial component in root canal disinfection.