香莲外洗液逆转白念珠菌耐药的转录组学研究

Xianglian External Lotion Restored the Sensitivity of Drug-resistantC andida albicans Strains to Fluconazole: a Transcriptomics Study

目的对香莲外洗液诱导白念珠菌标准耐药菌株恢复对氟康唑敏感前后的差异基因进行转录组学研究。方法采用微量肉汤稀释法测定白念珠菌标准耐药菌株最低抑菌浓度值(minimal inhibitory concentration,MIC),观察香莲外洗液诱导下白念珠菌标准耐药菌株恢复对氟康唑敏感所需的代数;通过转录组测序(RNA-Seq)测定和比较原代耐药株、香莲外洗液诱导下恢复敏感株的转录组情况;应用基因本体(gene ontology,GO)方法分析香莲外洗液诱导下恢复敏感菌株的差异表达基因。结果香莲外洗液诱导下的耐药白念珠菌转种至第6代时恢复对氟康唑的敏感性。转录组测序表明,对比原代耐药株,香莲外洗液诱导下恢复敏感株有165个基因RPKM(reads per kilo bases per million reads)指标表达上调,144个基因RPKM指标表达下调。GO分析发现:上下调基因主要归类于GO:0015903(fluconazole transport)。结论香莲外洗液可诱导耐药白念珠菌恢复对氟康唑的敏感性,推测可能与开启氟康唑转运通路有关。

Objective To perform a transcriptomics study in differential genes after Xianglian External Lotion（ XEL） induced the recovery of drug-resistant Candida albicans strains sensitive to Fluconazole. Methods Broth microdilution antifungal susceptibility test was used to detect minimal inhibitory concentration（ MIC） of drug-resistant Candida albicans strains induced by XEL. Transcriptome sequencing（ RNA-Seq） was used to determine and compare the transcription of primary drug-resistant Candida albicans strains and sensitive strains induced by XEL. High expressed genes and signaling pathways strains were analyzed by gene ontology（ GO） method. Results XEL could induce drug-resistant strains of the 6th generations to recover sensitivity. Transcriptome sequencing showed that,as compared with primary drug-resistant strains,there were 165 genes with up-regulated RPKM index and 144 genes with down-regulated RPKM index after XEL induction.GO analyses found that all genes were mainly classified as GO： 0015903（ fluconazole transport）. Conclusions XEL could induce the recovery of drug-resistant Candida albicans strains sensitive to Fluconazole. By analyzing transcriptomes,authors speculated that XEL could recover strain sensitivity to fluconazole by opening fluconazole transport pathway.