大蒜素联合环丙沙星抑制表皮葡萄球菌生物被膜的实验研究

Research on the Antibacterial Activity of Allicin Combined with Ciprofloxacin in Epidermal Staphylococcus Biofilm

目的:探索临床治疗细菌生物被膜感染的新途径,减少抗生素使用,降低细菌耐药率。方法:利用试管二倍稀释法检测大蒜素、环丙沙星及两者合用时抑制表皮葡萄球菌生物被膜时各自的MIC值;另一方面,将环丙沙星组、大蒜素+环丙沙星组药物作用后,观察生物被膜的生长曲线;同时,采用QT-PCR技术,探讨两组药物作用后生物被膜相关基因icaA表达的变化。结果:单用抑制表皮葡萄球菌被膜时,MIC大蒜素为(28.125±3.12)μg/mL,MIC环丙沙星为(3.125±0.23)μg/mL;两者联用时,MIC大蒜素、MIC环丙沙星分别为(14.063±2.30)μg/mL、(1.563±0.09)μg/mL。两组数据采用t检验分析后发现联用前后MIC有显著差异(P<0.05),且明显降低;大蒜素+环丙沙星组药物作用后,表皮葡萄球菌被膜生长抑制程度增强,且基因icaA的表达明显下降。结论:大蒜素联用后,可极大减少抗生素的用量、抑制表皮葡萄球菌被膜生长、降低被膜相关基因的表达,具有较高的体外抗菌活性。其不仅可为研究中药对细菌生物被膜的抑制及临床治疗相关细菌生物被膜感染提供坚实的实验数据参考,也将为降低临床细菌耐药率提供新的方向。

Objective:To explore a new way to treat bacterial biofilm infection to reduce the use of antibiotics and bacterial resistance.Methods:Respective MIC was measuring when allicin or ciprofloxacin was used on Epidermal Staphylococcus Biofilm by tube double dilution method.The bacterial growth curves were observed after the use of ciprofloxacin or allicin with ciprofloxacin.Meanwhile,the author explored the changes of icaA expression after drug action by QT-PCR.Results:MIC of allicin was(28.125±3.12)μg/mL and that of ciprofloxacin was(3.125±0.23)μg/mL when they inhibited Epidermal Staphylococcus Biofilm alone.The respective MIC was(14.063±2.30)μg/mL and(1.563±0.09)μg/mL when they combined treatment on Epidermal Staphylococcus Biofilm.There were significant differences between two sets of MIC by test analysis(P<0.05)and both of MIC declined.The bacterial growth inhibition enhanced but the expression of icaA decreased significantly after combination.Conclusion:Allicin combined with ciprofloxacin has high antibacterial activity,which can greatly reduce the dosage of antibioticsand inhibit the growth of Epidermal Staphylococcus Biofilm and reduce the expression of icaA.It not only can provide reliable expeimental basis for traditional Chinese medicine on inhibition of bacterial biofilm activity and clinical treatment of bacterial biofilm infections,it also can offer a direction for reducing clinical bacterial resistance.