汉防己甲素对氟康唑抗白念珠菌生物膜增效活性的初步研究

摘要

目的 探讨汉防己甲素( tetrandrine,TET)对氟康唑(fluconazole,FLC)抗白念珠菌生物膜是否有增效活性.方法 构建白念珠菌生物膜,参照微量稀释法,测定FLC单独及其联合TET对生物膜不同时期的最小抑菌浓度( minimum inhibitory concentration,MIC)；生物膜重新悬浮后,测定FLC单独及其联合TET对不同浓度菌液的MIC.结果 FLC单独及其联合TET对白念珠菌生物膜最初期(0 h)的MIC50值范围分别为0.25～64μg/mL和0.125～16 μg/mL(P=0.002)；早期(4 h)的MIC50值范围分别为8～256μg/mL和1～64 μg/mL(P=0.000)；中期(24 h)、成熟期(48 h)的MIC50值均＞1024 μg/mL.生物膜重新悬浮后,FLC单独及其联合TET对低浓度菌液(终浓度为1×103 CFU/mL)的MIC值范围分别为0.25～64μg/mL和0.125～16 μg/mL(P=0.003),高浓度菌液(终浓度为1×106 CFU/mL)的MIC值均＞64 μg/mL.结论 汉防已甲素在体外对氟康唑抗白念珠菌生物膜最初期(0 h)、早期(4 h)有增效活性,对中期(24 h)、成熟期(48 h)无增效活性；汉防己甲素对氟康唑抗白念珠菌生物膜重新悬浮后的低浓度菌液(终浓度为1×103 CFU/mL)有增效活性,高浓度菌液(终浓度为1×106 CFU/mL)无增效活性.%The aim was to investigate the synergistic effect of tetrandrine (TET) on fluconazole (FLO to Candida albi-cans biofilms. The biofilms of C. albicans was built, and the minimum inhibitory concentration (MIC) of FLC alone as well as in combination with TET on each stage of biofilms forming were determined by the CLSI proposed M27-A microdilution method. The MIC of FLC alone and in combination with TET on re-suspended C. albicans biofilms were tested. In initial stage (Oh), the MIC50 of FLC alone and combined with TET to the C. albicans biofilms were 0. 25-64 g/mL and 0. 125-16 g/mL respectively (P=0. 002). In the early stage (4thh), the MIC50 were 8-256μg/mL and 1-64 μg/mL (P = 0. 000). In the middle (24lhh) and advanced stages (48th h), both the MIC50 were >1024 μg/mL. The MIC of FLC alone to the re-suspended C. albicans biofilm (1×103 CFU/mL) were 0. 25-64 μg/mL and 0. 125-16 μg/mL when combined with TET. Both the MIC of FLC alone and in combination with TET were >64 μg/mL in the cells counts of 1×10s CFU/mL. In vitro, TET acts synergistical-ly with fluconazole in the activity against the C. albicans biofilms in initial stage (Oh) and early stage (4thh) , without synergistic effect in middle stage (24lhh) and advanced stage (48th h) ; there is synergistic effect of tetrandrine to fluconazole against the re-suspended thalli of C. albicans biofilm (1×103CFU/mL) ,and no synergy against the thalli (1×106CFU/mL).