铁离子在耳念珠菌热抗逆中功能及机制初探研究  

A preliminary study on the effect of iron on heat resistance mechanism in Candida auris

作　　者：文欢 王也 黄雎 周浩 王棣[1] 宋章永 WEN Huan;Wang Ye;HUANG Ju;ZHOU Hao;WANG Di;SONG Zhangyong(Department of Medical Laboratory,Xuyong County People’s Hospital,Luzhou 646400,China;School of Basic Medical Sciences,Southwest Medical University,Luzhou 646000,China)

机构地区：[1]四川省泸州市叙永县人民医院检验科,泸州646400 [2]西南医科大学基础医学院,泸州646000

出　　处：《中国真菌学杂志》2025年第1期45-51,共7页Chinese Journal of Mycology

基　　金：四川省自然科学基金项目(2025ZNSFSC0602);泸州市-西南医科大学联合项目(2023LZXNYDJ031);叙永县人民医院-西南医科大学科技战略合作项目(2023XYXNYD08);临床疾病分子诊断泸州市重点实验室科技项目(FZZD2023-07)。

摘　　要：目的探究铁离子在耳念珠菌热抗逆中的生物学功能及其机制。方法首先,采用液体生长曲线方法测定不同温度和不同浓度铁螯合剂处理对耳念珠菌和希木龙念珠菌菌株生长的影响。其次,采用固体点板试验测定两株菌株在不同温度、不同浓度铁离子处理下的生长变化。再次,通过DCFH-DA荧光探针方法测定耳念珠菌菌株在不同温度下、不同浓度铁离子处理下菌体内活性氧(reactive oxygen species,ROS)的变化。最后,采用荧光定量PCR方法测定耳念珠菌菌株的单巯基谷氧化还原蛋白基因(GRX5)在不同温度、不同浓度铁离子和加入H_(2)O_(2)情况下的表达量变化。结果耳念珠菌菌株比希木龙念珠菌株表现出更强的耐热特性。高温条件下,缺铁显著抑制耳念珠菌菌株生长,富集浓度铁显著促进耳念珠菌菌株生长。添加铁螯合剂处理抑制耳念珠菌菌株产生ROS,而富集浓度铁则促进耳念珠菌菌株产生ROS,且不同温度、不同浓度铁离子和H_(2)O_(2)条件下GRX5基因表达量存在差异。结论铁离子是耳念珠菌热抗逆的必需因素,且通过GRX5基因表达上调来抵御热胁迫带来的ROS压力。Objective To explore the biological effect and molecular mechanism of iorn in heat resistance of Candida auris isolate.Methods Firstly,the OD_(600)growth curves of C.auris and C.hemulosaurus strains were measured under different temperatures and concentrations of iron chelator bathophenanthroline disulfonate in liq-uid medium treatment.Secondly,solid spot plate experiments at different temperatures and con-centrations of iron were performedin phenotypic analysis of C.auris and C.hemulosaurus strains.Thirdly,DCFH-DA fluorescent probe was used to determine the production of reactive oxygen species(ROS)in C.auris strain under different temperatures and concentrations of iron.Fourth-ly,the changes of glutaredoxin 5 gene(GRX5)expression under the conditions of different tem-peratures,concentrations of iron or H_(2)O(2) addition were detected by real-time PCR method.Results Compared with the C.hemulosaurus strain,C.auris strain showed stronger heat tolerance.The growth of C.auris strain was significantly promoted in the solid medium enriched with high con-centration of iron.In addition,the addition of iron chelator bathophenanthroline disulfonate would inhibit the ROS production of C.auris strain,while the enrichment of iron would promote the ROS production.Further more,real-time PCR analysis showed that the expression of GRX5 gene of C.auris strain was changed under different temperatures and concentrations of iron.Conclusion Iron is a neces-sary factor for the thermal stress resistance of C.auris strain.Moreover,the expression GRX5 gene is up-regulated to resist ROS stress caused by heat stress.

关 键 词：耳念珠菌 希木龙念珠菌 铁离子 热抗逆 单巯基谷氧化还原蛋白基因 

分 类 号：R379.4[医药卫生—病原生物学]