中药小檗碱协同抗真菌药对马尔尼菲篮状菌的体外药敏试验研究

In vitro susceptibility of berberine combined with antifungal agents against Talaromyces marneffei

目的探讨小檗碱单独及与常规抗真菌药物联合使用后对马尔尼菲篮状菌酵母相的作用效果。方法采用CLSI棋盘法对21株马尔尼菲篮状菌(Talaromyces marneffei,TM)进行体外药敏试验,使用分数抑制浓度指数(FICI)判断药物相互作用:FICI≤0.5为协同作用,FICI>4为拮抗作用,0.5<FICI≤4为无关作用。利用时间-杀菌曲线进一步确认相互作用。结果小檗碱联合常用抗真菌药物具有协同作用,协同作用率分别为:小檗碱/氟康唑(47.62%),小檗碱/伊曲康唑(66.67%),小檗碱/伏立康唑(71.43%),小檗碱/两性霉素B(71.43%),小檗碱/卡泊芬净(52.38%)。时间-杀菌实验结果显示,5组联合用药组在24 h时每毫升TM的CFU的对数值均与任意单用药物组相比下降大于等于2个单位。所有组合均未观察到拮抗作用。结论中药小檗碱体外对马尔尼菲篮状菌具有抑制作用,与常规抗真菌药联合使用可大大降低抗真菌药物对马尔尼菲篮状菌酵母相的MIC值。这结论为小檗碱联合抗真菌药应用于马尔尼菲篮状菌病的治疗提供实验室基础。

Objective To investigate the effect of berberine alone or in combination with conventional antifungal drugs on Talaromyces marneffei yeast.Methods Twenty-one isolates of T.marneffei were assayed.Drug interactions were assessed with the chequerboard technique using the CLSI microdilution method(M27-A3)with minor modifications.The fractional inhibitory concentration index(FICI)was used to classify drug interactions.Time-kill curves were used to confirm the synergistic interaction.Results Berberine/voriconazole and berberine/amphotericin B combination showed the highest percentage of synergic interaction(71.4%);followed by berberine/itraconazole(66.67%),berberine/fluconazole(47.62%),berberine/caspofungin(52.38%),respectively.Time-kill curves confirmed the synergistic interaction,no antagonistic or indifferent action was observed.Conclusions In vitro berberine could enhances the efficacy of antifungal agents against the yeast form of T.marneffei.The MIC value can be greatly reduced when combined with conventional antifungal drugs.The results indicated berberine might have a potential role in combination with antifungal agents in freating patients infected with T.marneffei.