目的 探讨黄连解毒汤醋酸乙酯提取物(ethyl acetate extract of Huanglianjiedu Decoction,EAHJD)对白色念珠菌毒力因子的作用.方法 分别采用卵黄培养基法检测磷脂酶(PL)活力、牛奶平板法检测天冬氨酸蛋白酶(Sap)活力、橄榄油乳化法检测脂肪酶(Lip)活力;水-烃两项测定实验检测细胞表面疏水性(CSH);RT-PCR法检测毒力因子相关基因的表达.结果 EAHJD对PL的活力无影响;1 250 μg/mL EAHJD可显著抑制Sap与Lip的活力,312 μg/mL EAHJD效果次之;EAHJD对CSH呈现剂量依赖性抑制,CSH1分别下调了7.69、3.57、2.95倍;分泌型酶相关基因在EAHJD作用下呈现不同倍数的变化,其中PLC1、Sap2、Sap3、Sap9、Lip3、Lip4、Lip6下调,PLB1、PLC2、Sap10、Lip5无明显变化.结论 EAHJD可抑制白色念珠菌毒力因子的活力.

Objective To observe the effects of ethyl acetate extract from Huanglian Jiedu Decoction (EAHJD) on the virulence factors of Candida albicans. Methods Egg-yolk medium, milk-plate medium, and olive oil emulsification were used respectively to test the activities of phospholipase (PL), aspartic protease (Sap), and lipase (Lip) of C. albicans. The water-hydrocarbon two-phase assay was applied to measure the cell surface hydrophobicity (CSH) of C. albicans. qRT-PCR was adopted to observe the expression of virulence factors related genes. Results EAHJD had no effect on the activity of PL. EAHJD (1 250 μg/mL) could significantly inhibit the activity of PL and Lip better than that by 312 μg/mL EAHJD. EAHJD could reduce CSH of C. albicans in a dose-independent manner and CSH1 was down-regulated by 7.69, 3.57, and 2.95 folds by 1 250, 312, and 78 μg/mL EAHJD, respectively. The expression of secretory enzyme related genes displayed different changing folds: PLC1, Sap2, Sap3, Sap9, Lip3, Lip4, and Lip6 were down-regulated; PLB1, PLC2, Sap1, Sap10, and Lip5 had no distinct change treated by EAHJD. Conclusion EAHJD could inhibit the activities of virulence factors of C. albicans.

国家自然科学基金资助项目(81073127);安徽省自然科学基金项目(1408085MH165);安徽中医药大学校级科研项目(2014zr026)