黄芩苷联合氟康唑通过降低 cAMP 抑制白念珠菌酵母-菌丝形态转化

摘要

目的：探讨中药有效成分黄芩苷(Baicalin,BA)联合氟康唑(Fluconazole,FLC)对白念珠菌(Candida albicans )形态转化的影响及机制。方法在菌丝诱导培养基(液体和固体培养基)上观察两药联用对白念珠菌酵母-菌丝形态转化的影响；ELISA 法检测两药联用对胞内 cAMP 含量的影响；回补实验验证两药联用对胞内 cAMP 的影响；qRT-PCR 检测两药联用对白念珠菌 cAMP 相关基因 RAS1,CDC35和 PDE2表达的影响。结果黄芩苷联合氟康唑能显著抑制白念珠菌酵母-菌丝形态转化,降低胞内 cAMP 水平；外源性 cAMP 能逆转两药联用所造成的菌丝抑制；两药联用使 RAS1和 CDC35表达分别下调35%和27%,使 PDE2上调1.21倍。结论黄芩苷协同氟康唑显著抑制白念珠菌形态转化,其作用机制可能与下调胞内 cAMP 有关。%Objective This study aimed to investigate the antifungal activity of baicalin in combination with fluconazole a-gainst Candida albicans morphological transition from yeast to hyphae and to explore the mechanism.Methods The hypha-inducing liquid and solid media were uesd to observe the effect of baicalin alone or in combination with fluconazole on yeast-hapal transformation in C .albicans ;The intracellular cAMP level and cAMP rescue was measured by ELISA Kit;Quantitative real time PCR (qRT-PCR)was performed to measure the expression of cAMP-related genes.Results Baicalin in combination with fluconazole could inhibit hyphae formation and intracellular cAMP level obviously;exogenous cAMP could convert hy-pha-inhibiting effect;And expression of RAS1 and CDC35 were downregulated by 35% and 27% respectively,while the ex-pression of PDE2 was upregulated by 121%.Conclusion Mechanism of baicalin in combination with fluconazole inhibiting C . albicans morphological transition was probabaly via down-regulating the intracellular cAMP.