盐酸氨溴索对体外白色假丝酵母菌成熟生物膜的抑制作用及其形态学的影响

Inhibition of ambroxol hydrochloride on Candida albicans biofilm in vitro and its effect on morphogenesis

目的研究盐酸氨溴索对体外白色假丝酵母菌(Candida albicans)成熟生物膜(biofilm,BF)的影响。方法用微孔板法建立体外白色假丝酵母菌ATCC 90028 BF模型;采用甲基四氮盐(the abated tetrazolium salt,XTT)减低法定量评价盐酸氨溴索对白色假丝酵母菌成熟BF的抑制作用;银染后,倒置显微镜下观察该药对白色假丝酵母菌成熟BF的形态学影响。结果在96孔微量细胞培养板上成功建立白色假丝酵母菌BF模型;1.25、2.5、5、7.5 mg/ml的盐酸氨溴索作用白色假丝酵母菌成熟BF 12 h后,XTT减低法D(450)值分别为(0.63±0.05)、(0.52±0.08)、(0.31±0.05)和(0.11±0.03),分别与空白对照组(0.71±0.07)比较,差异有显著性(P<0.05);0.625 mg/ml的盐酸氨溴索作用白色假丝酵母菌成熟BF后,D(450)值为(0.67±0.04),与空白对照组比较,差异无显著性(P>0.05);不同浓度的盐酸氨溴索作用白色假丝酵母菌成熟BF,组间比较,均有显著性差异(P<0.05)。结论盐酸氨溴索对体外白色假丝酵母菌成熟BF有抑制作用,且在一定药物浓度范围内随着该药药物浓度的增加,对BF的抑制作用显著增强。

Objective To investigate the effect of ambroxol hydrochloride on biofilm and morphology of Candida albicans（C.albicans） in vitro.Methods Microtiter plate culture method was used to establish C.albicans（ATCC 90028） biofilm.The abated tetrazolium salt（XTT） reduction assay was performed to quantitatively analyze the effect of ambroxol hydrochloride against C.albicans mature biofilm for 12 h.Microscopic examination with silver staining was conducted to assess the effect of ambroxol hydrochloride on morpho-logy of C.albicans biofilm.Results C.albicans biofilm in vitro was formed on 96-well microtiter plates successfully.XTT reduction assay indicated that the OD450 values at different concentration of ambroxol hydrochloride（0.625,1.25 and 2.50,5.00 and 7.50 mg/ml） were 0.68±0.04,0.63±0.05,0.52±0.07,0.31±0.05 and 0.11±0.03,respectively.All of these values had significant difference when compared with that of control（0.71±0.07,P0.05）.But,when the concentration of ambroxol hydrochloride was 0.625 mg/ml,the OD450 value was 0.67±0.04,which had no significant difference with that of control（P0.05）.After the treatment of ambroxol hydrochloride（2.50,5.00 and 7.50 mg/ml） for 48 h,biofilm structure was destroyed,extracellular matrix was decreased,more single yeast cells were observed.Conclusion Ambroxol hydrochloride inhibits mature biofilm of C.albicans in a dose-dependent manner.