亚抑菌浓度氟康唑对光滑念珠菌Sap活性和生物膜形成的影响  

The effect of sub minimum inhibitory concentration fluconazole on Sap and biofilm formation of Candida glabrata

作　　者：程鹏 张倩[2] 徐雯 马继登[2] 巢世兰 杨春娇[2] 阿祥仁 CHENG Peng;ZHANG Qian;XU Wen;MA Jideng;CHAO Shilan;YANG Chunjiao;A Xiangren(Department of Basic Medicine,College of Medical,Qinghai University,Xining,Qinghai 810016,China;Department of Medical Laboratory,Qinghai Provincial People's Hospital,Xining,Qinghai 810016,China)

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

出　　处：《热带医学杂志》2024年第8期1078-1083,共6页Journal of Tropical Medicine

基　　金：青海省2022年创新平台建设专项项目(2022-ZJ-T01)。

摘　　要：目的探究亚抑菌浓度(sub-MIC)氟康唑对光滑念珠菌分泌的天冬氨酰蛋白酶(Sap)活性和生物膜形成的影响,为临床诊疗提供理论依据。方法收集青海省人民医院2021年4月-2022年10月临床分离的光滑念珠菌,采用半自动细菌分析系统法测定氟康唑对光滑念珠菌的最低抑菌浓度(MIC)。采用牛血清蛋白琼脂培养基检测sub-MIC氟康唑对光滑念珠菌Sap活性的影响;结晶紫法检测sub-MIC氟康唑对光滑念珠菌生物膜形成的影响;实时荧光定量逆转录聚合酶链式反应(RT-qPCR)法检测在不同sub-MIC氟康唑作用下光滑念珠菌Sap和生物膜形成相关基因转录水平,分为对照组(不含氟康唑)、1/2MIC氟康唑组、1/4MIC氟康唑组、1/8MIC氟康唑组和1/16MIC氟康唑组。结果氟康唑对9株临床分离的光滑念珠菌MIC为8~32 mg/L。与对照组相比,1/2MIC氟康唑组光滑念珠菌Sap活性明显升高,差异有统计学意义(F=5.917,P<0.05),而1/4MIC、1/8MIC和1/16MIC氟康唑各组Sap活性差异均无统计学意义(P均>0.05);1/2MIC和1/4MIC氟康唑可明显刺激YPS3、YPS5和YPS8基因转录(F=4.271、7.551、2.945,P均<0.05),1/8MIC和1/16MIC氟康唑对Sap相关基因转录无明显干预作用(P>0.05)。与对照组相比,不同sub-MIC氟康唑对光滑念珠菌生物膜形成无明显干预作用,差异无统计学意义(P>0.05),各组SNF2、SNF6、EFG1、TEC1和YAK1基因转录水平差异均无统计学意义(P均>0.05);与对照组相比,1/2MIC氟康唑可下调CST6基因转录,差异有统计学意义(F=4.002,P<0.05),1/4MIC、1/8MIC和1/16MIC氟康唑组CST6基因转录差异均无统计学意义(P均>0.05)。结论1/2MIC氟康唑可能上调YPS3、YPS5和YPS8基因转录水平,从而促进光滑念珠菌分泌Sap。sub-MIC氟康唑对光滑念珠菌生物膜形成无明显作用。Objective To investigate the effects of sub minimum inhibitory concentration(sub⁃MIC)of fluconazole on the secreted aspartyl protease(Sap)and biofilm formation of Candida glabrata.Methods Clinical isolates of C.glabrata were collected from the Qinghai Provincial People's Hospital from April 2021 to October 2022;the minimum inhibitory concentration(MIC)of fluconazole against C.glabrata was determined by automatic testing bacteriology method.The effect of sub⁃MIC fluconazole on Sap activity of C.glabrata was detected by bovine serum protein agar medium.The effect of sub⁃MIC fluconazole on the biofilm formation of C.glabrata was detected by crystal violet method.Reverse transcription⁃quantitative polymerase chain reaction(RT⁃qPCR)was used to detect the transcription levels of Sap and biofilm formation related genes in C.glabrata treated with different sub⁃MIC of fluconazole.It was divided into control group(without fluconazole),1/2MIC fluconazole group,1/4MIC fluconazole group,1/8MIC fluconazole group and 1/16MIC fluconazole group.Results The MIC of fluconazole against 9 clinical isolates of C.glabrata was 8-32 mg/L.Compared with the control group,the Sap activity of C.glabrata was significantly increased in the 1/2MIC fluconazole group(F=5.917,P<0.05),while there was no significant difference in the 1/4MIC,1/8MIC and 1/16MIC fluconazole groups(all P>0.05).Fluconazole at 1/2MIC and 1/4MIC significantly stimulated the transcription of YPS3,YPS5 and YPS8 genes(F=4.271,7.551,2.945;all P<0.05),while fluconazole at 1/8MIC and 1/16MIC had no significant effect on the transcription of YPS3,YPS5 and YPS8 genes(P>0.05).Compared with the control group,different sub⁃MICs of fluconazole had no significant effect on the biofilm formation of C.glabrata(P>0.05),and there were no significant differences in the transcription levels of SNF2,SNF6,EFG1,TEC1 and YAK1 genes among the groups(all P>0.05).The CST6 gene expression was down⁃regulated by 1/2MIC fluconazole,compared with the control group,the difference was stat

关 键 词：光滑念珠菌 亚抑菌浓度 氟康唑 天冬氨酰蛋白酶 生物膜 

分 类 号：R379.4[医药卫生—病原生物学]