牡丹皮乙醇提取物对热带念珠菌毒力因子的影响  

The effect of ethyl alcohol extract of moutan cortex on virulence factors of Candida tropicalis

作　　者：唐赟 阿祥仁[2] 程鹏[2] 周健武 马继登 马小亚 Tang Yun;A Xiangren;Cheng Peng;Zhou Jianwu;Ma Jideng;Ma Xiaoya(Department of Basic Medicine,College of Medicine,Qinghai University,Xining 810016,China;Department of Medical Laboratory,Qinghai Provincial People′s Hospital,Xining 810007,China)

机构地区：[1]青海大学医学院基础医学部,西宁810016 [2]青海省人民医院医学检验科,西宁810007

出　　处：《中华检验医学杂志》2025年第2期258-264,共7页Chinese Journal of Laboratory Medicine

基　　金：青海省2022年创新平台建设专项(2022-ZJ-T01)。

摘　　要：目的探究牡丹皮乙醇提取物(EAEMC)对热带念珠菌的体外抑菌效果,以及对毒力因子包括天冬氨酸蛋白酶、溶血素、磷脂酶、酯酶、脂肪酶的活性和生物膜的影响。方法通过超声提取、减压浓缩和冻干得到EAEMC粉末;以微量稀释法测定EAEMC对21株热带念珠菌临床菌株和1株标准菌株的最低抑菌浓度(MIC)和最低杀菌浓度(MBC);通过平板法检测热带念珠菌的5种胞外酶活性以及EAEMC对其影响,结果采用方差分析;体外构建热带念珠菌生物膜模型,噻唑蓝法评价EAEMC对热带念珠菌生物膜抑制率。结果EAEMC对热带念珠菌BNCC335988的MIC为12.5 g/L,MBC为25 g/L,对临床菌株的MIC为12.5~25 g/L,MBC为25~50 g/L;热带念珠菌的天冬氨酸蛋白酶、酯酶和溶血素活性均为阳性,磷脂酶和脂肪酶均为阴性;EAEMC浓度为1/2MIC时能完全抑制热带念珠菌的天冬氨酸蛋白酶和溶血素活性,为MIC时能完全抑制酯酶活性;EAEMC浓度为2MIC时对热带念珠菌BNCC335988生物膜的抑制率达到70%以上,浓度为4MIC时达到80%以上,浓度为8MIC时抑制率达90%以上。结论EAEMC可通过降低热带念珠菌的天冬氨酸蛋白酶、酯酶和溶血素的活性,以及抑制生物膜的形成来达到抑菌作用。Objective To investigate the in vitro bacteriostatic effect of ethyl alcohol extract of Moutan Cortex(EAEMC)on Candida tropicalis and its effect on virulence factors,including aspartic protease,hemolysin,phospholipase,esterase,lipase activities and biofilm.MethodsEAEMC powder was obtained by ultrasonic extraction,decompression concentration and lyophilization;the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of EAEMC on 21 clinical strains and one standard strain of Candida tropicalis were determined by microdilution.Five extracellular enzyme activities of Candida tropicalis and the effect of EAEMC on them were detected by the plate assay,and the results were analyzed by ANOVA.The biofilm model of Candida tropicalis was constructed in vitro,and the inhibition rate of EAEMC on Candida tropicalis biofilm was evaluated using the thiazolyl blue(MTT)method.ResultsThe MIC of EAEMC against Candida tropicalis BNCC335988 was 12.5 g/L and the MBC value was 25 g/L,while for the clinical strains,the MIC was 12.5-25 g/L and the MBC was 25-50 g/L.Aspartic protease,esterase and hemolytic activities of Candida tropicalis were positive,but phospholipase and lipase showed negative activities.At a concentration of 1/2 MIC of EAEMC,the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited the aspartic protease and hemolytic activities of Candida tropicalis were completely inhibited and the esterase activity was completely inhibited at a concentration of MIC of EAEMC.The inhibition of Candida tropicalis BNCC335988 biofilm by EAEMC reached more than 70%at a concentration of 2MIC,more than 80%at a concentration of 4MIC,and more than 90%at a concentration of 8MIC.ConclusionEAEMC can achieve bacteriostatic effects by reducing the aspartic protease,esterase and hemolysin activities of Candida tropicalis,as well as inhibiting biofilm formation.

关 键 词：念珠菌 热带 牡丹皮乙醇提取物 毒力因子 胞外酶 生物膜 

分 类 号：R285[医药卫生—中药学]