芍药汤对白念珠菌生物膜体外抑制活性研究

Study on the In Vitro Inhibitory Activity of Shaoyao Decoction on Candida Albicans Biofilms

目的:观察经方芍药汤对白念珠菌生物膜的体外抑菌效果。方法:应用酵母菌肉汤稀释抗真菌药敏试验方案,测定芍药汤对白念珠菌最小抑菌浓度(MIC_(50))和最小杀菌浓度(MBC),运用扫描电子显微镜(SEM)及激光共聚焦显微镜(CLSM)观察芍药汤干预白念珠菌作用时的形态变化,并采用甲基四氮盐(XTT)减低法及结晶紫染色法测定芍药汤对白念珠菌生物膜的最小抑制浓度(SMIC_(50))、代谢活性及生物膜量的影响。结果:芍药汤对白念珠菌的MIC_(50)和MBC为1.60 mg/mL和20.15 mg/mL;芍药汤对白念珠菌生物膜的SMIC_(50)为5.04 mg/mL;芍药汤可破坏白念珠菌生物膜结构及菌丝的生长;在不同时间点(2、6、12、24、48 h)实验组与空白对照组比较,芍药汤对白念珠菌生物膜生成活性具有抑制作用(P<0.05),然而在2 h时,实验组与空白对照组比较,差异无统计学意义(P>0.05),6、12、24、48 h时间点生物膜生成活性均显著增加(P<0.05),但24 h生物膜生成量相比于48 h生物膜生成量,差异无统计学意义(P>0.05)。在2~12 h白念珠菌生物膜的整体代谢活性呈下降趋势(P<0.05),然而在12~48 h白念珠菌整体代谢活力性比较,差异无统计学意义(P>0.05),XTT减低法和结晶紫染色测定法对白念珠菌生物膜测定的结果,差异无统计学意义(P>0.05)。结论:芍药汤对白念珠菌具有良好的体外抑菌活性,芍药汤抑制白念珠菌生物膜具有浓度依赖性,并通过延长芍药汤的药物作用时间达到抑制白念珠菌生物膜的效果。

Objective:To investigate the in vitro antibacterial effect of Shaoyao Decoction on Candidia.albicans.Methods:Using the yeast broth dilution antifungal susceptibility test protocol,the minimum inhibitory concentration(MIC_(50))and minimum bactericidal concentration(MBC)of Shaoyao Decoction against C.albicans were determined.Scanning electron microscopy(SEM)and laser scanning confocal microscopy(LSCM)were employed to observe morphological changes in C.albicans upon treatment with the decoction.Additionally,the Methyl Tetrazolium Salt(XTT)reduction assay and crystal violet staining method were used to assess the minimum suppression concentration(SMIC_(50)),metabolic activity,and biofilm mass of C.albicans biofilms treated with the decoction.Results:The MIC_(50) and MBC of Shaoyao Decoction for C.albicans were found to be 1.60 mg/mL and 20.15 mg/mL,respectively,with an SMIC_(50) for C.albicans biofilms at 5.04 mg/mL.Paeoniae decoction disrupts the structure and hyphal growth of C.albicans biofilm.Comparisons between the experimental group and the blank control group at different time points(6,12,24,and 48 hours)showed that Paeoniae decoction significantly inhibited the biofilm formation activity of C.albicans(P<0.05).However,at 2 hours,there was no statistically significant difference between the experimental group and the blank control group(P>0.05).At 6,12,24,and 48 hours,the biofilm formation activity significantly increased(P<0.05),but the difference in biofilm formation between 24 hours and 48 hours was not statistically significant(P>0.05).The overall metabolic activity of C.albicans biofilm showed a decreasing trend from 2 to 12 hours(P<0.05).However,from 12 to 48 hours,the overall metabolic activity of C.albicans showed no statistically significant difference(P>0.05).The results of biofilm determination by XTT reduction assay and crystal violet staining method showed no statistically significant differences(P>0.05).Conclusion:Shaoyao Decoction exhibits effective in vitro antifungal activity against C.albicans,with a concentration-dependent inhibition of C.albicans biofilms.Prolonging the duration of treatment with the decoction can enhance its inhibitory effect on C.albicans biofilms.