摘要
目的探讨丹参酮单体与β-内酰胺类抗菌药物联用对表皮葡萄球菌(Staphylococcus epidermidis,SE)早期生物膜形成的抑制作用及其机制。方法采用半定量黏附实验检测SE生物膜基质含量、XTT实验检测膜内菌代谢活性,以及qRT-PCR检测生物膜形成关键基因,来验证丹参酮单体与β-内酰胺类抗菌药物联用的药效作用及可能的机制。结果 2μg/ml的二氢丹参酮Ⅰ和2μg/ml的氨苄西林联用对SE生物膜具有协同抑制作用(FICI<0.5);两药联用可明显抑制SE黏附(6h)、聚集阶段(24h)基质形成,影响生物膜内细菌代谢活性并杀死细菌,且联用组优于2μg/ml二氢丹参酮Ⅰ、2μg/ml氨苄西林单药组;联用组降低了黏附阶段SE生物膜形成关键基因icaA、atlE、aap和stk的表达量,但并不能抑制聚集阶段上述基因的表达。结论二氢丹参酮Ⅰ和氨苄西林联用对SE生物膜形成早期具有协同抑制作用,其机制可能与影响基质形成、细胞代谢活性以及生物膜形成关键基因的表达有关。
OBJECTIVE To observe the inhibitory effect and mechanisms of tanshinone monomer combined withβ-lactams antibiotics on early formation of Staphylococcus epidermidis biofilms.METHODS S.epidermidi biofilm matrix contents were detected with the use of semi-quantitative adhesion experiment,the metabolic activity was determined by means of XTT assay,and the key genes for biofilm formation were detected by using qRT-PCR so as to verify the therapeutic effect of tanshinone monomer combined withβ-lactams antibiotics and the possible mechanisms.RESULTS 2μg/ml of dihydrodanshinoneⅠ combined with 2μg/ml of ampicillin had synergistic and inhibitory effect on S.epidermidis biofilms（FICI0.5）;the combined use of two drugs had remarkable inhibitory effect on adhesion of S.epidermidis（6 h）,formation of matrix at accumulation phase（24 h）,affected the bacterial metabolic activity and killed bacteria.The therapeutic effect of the combined use of the drugs was better than that of the single use of 2μg/ml of dihydrodanshoneoneⅠ or 2μg/ml of ampicillin（P〈0.05）.The combined use of the drugs reduced the levels of expression of key genes for formation of the S.epidermidis biofilms such as icaA,atlE,aap and stk,but it could not inhibit the expression of the above genes at the accumulation phase.CONCLUSIONDihydrodanshinoneⅠ combined with ampicillin has the synergistic and inhibitory effect on early formation of S.epidermidis biofilms,and the mechanisms may affect the formation of matrix,bacterial metabolic activity and expression of the key genes for the formation of biofilms.
作者
雍江堰
祖瑞铃
易玉玲
李燕
YONG Jiang-yan;ZU Rui-ling;YI Yu-ling;LI Yan(College of Medical Technology, Chengdu University of TCM, Chengdu, Sichuan 611137, China)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2018年第7期966-970,982,共6页
Chinese Journal of Nosocomiology
基金
四川省科技厅应用基础项目(2015JY0159)
关键词
表皮葡萄球菌
生物膜
二氢丹参酮Ⅰ
氨苄西林
联合用药
Staphylococcus epidermidis
Biofilm
Dihydrodanshinone I
Ampicillin
Combined use of drugs
