高通量测序分析不同浓度的白鲜碱对须癣毛癣菌转录组的影响

Effect of dictamnine at different concentration on Trichophyton mentagrophytes transcriptome: based on high-throughput sequencing

摘要:

【背景】目前，由皮肤浅部真菌引起的皮肤病逐年增加，已经成为影响人类健康的重大问题之一。【目的】探究白鲜碱(dictamnine，DIC)单体对须癣毛癣菌的抑菌作用及作用机制。【方法】通过高通量测序技术对须癣毛癣菌及白鲜碱作用后的须癣毛癣菌进行转录组测序，对测序序列进行处理和生物信息学分析，以及测序质量评价和序列注释。通过与对照组比较，鉴别出差异表达基因，然后进行GO功能显著性分析和KEGG代谢途径分析等。【结果】白鲜碱对须癣毛癣菌的MIC值为50 μg/ml。DIC高剂量组与对照组比较，共360个差异基因；其中上调表达265个，下调表达95个。MFS1、KU70、KU80、L2、cpaT、MFS2、VdtG、patC等基因表达有显著差异。GO功能富集表明，差异基因主要集中于核糖体、线粒体膜、抗氧化活性、毒素代谢过程、单一生物代谢过程、次生代谢过程、初级活性跨膜转运蛋白活性和活跃的跨膜转运蛋白活性等。KEGG代谢途径的改变主要集中在ABC转运、膜运输障碍、谷胱甘肽代谢、DNA的复制与修复变化、减数分裂、氧化磷酸化和丙酮酸代谢障碍等富集通路上，并且相关基因均有显著性改变(P<0.05)。【结论】白鲜碱对须癣毛癣菌的抑菌机制可能与影响须癣毛癣菌DNA复制表达、能量代谢、DNA修复、ABC转运等途径有关。

Abstract:

[Background] Superficial mycosis cases have been increasing year by year and the pathogenic fungi have become one of the major threats to human health. [Objective] This paper aims to explore the inhibitory effect of dictamnine (DIC) on Trichophyton mentagrophytes and the underlying mechanism. [Methods] Transcriptome sequencing was performed on T. mentagrophytes and T. mentagrophytes after the action of DIC by high-throughput sequencing technology, processing and bioinformatics analysis of the sequencing sequence, sequencing quality evaluation and sequence annotation. By comparing with the control group, differentially expressed genes were identified, and then GO function significance analysis and KEGG metabolic pathway analysis were performed. [Results] The minimum inhibitory concentration (MIC) of DIC on T. mentagrophytes was 50 μg/mL. A total of 360 differentially expressed genes (DEGs) were identified between the control group and DIC high-dose group (265 up-regulated and 95 down-regulated), and the expression of MFS1, KU70, KU80, L2, cpaT, MFS2, VdtG and patC was significantly different. The DEGs were mainly involved in the GO terms of ribosome, mitochondrial membrane, antioxidant activity, toxin metabolic process, single biological metabolic process, secondary metabolic process, primary active transmembrane transporter activity, and active transmembrane transporter activity. The changes in the KEGG metabolic pathway mainly focus on enrichment pathways such as ABC transport, membrane transport barriers, glutathione metabolism, DNA replication and repair changes, meiosis, oxidative phosphorylation and pyruvate metabolism disorders, and related genes are all There is a significant change (P<0.05). [Conclusion] DIC inhibits T. mentagrophytes by influencing DNA replication, expression, and repair, energy metabolism, ABC transport, and other pathways of the fungus.