Xiao Chen-Wen, Ji Quan-An, Wei Qiang, Liu Yan, Pan Li-Jun, Bao Guo-Lian
Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou, Zhejiang Province, China.
PLoS One. 2015 Apr 14;10(4):e0124265. doi: 10.1371/journal.pone.0124265. eCollection 2015.
Berberine, a natural isoquinoline alkaloid of many medicinal herbs, has an active function against a variety of microbial infections including Microsporum canis (M. canis). However, the underlying mechanisms are poorly understood. To study the effect of berberine chloride on M. canis infection, a Digital Gene Expression (DGE) tag profiling was constructed and a transcriptome analysis of the M. canis cellular responses upon berberine treatment was performed. Illumina/Hisseq sequencing technique was used to generate the data of gene expression profile, and the following enrichment analysis of Gene Ontology (GO) and Pathway function were conducted based on the data of transcriptome. The results of DGE showed that there were 8476945, 14256722, 7708575, 5669955, 6565513 and 9303468 tags respectively, which was obtained from M. canis incubated with berberine or control DMSO. 8,783 genes were totally mapped, and 1,890 genes have shown significant changes between the two groups. 1,030 genes were up-regulated and 860 genes were down-regulated (P<0.05) in berberine treated group compared to the control group. Besides, twenty-three GO terms were identified by Gene Ontology functional enrichment analysis, such as calcium-transporting ATPase activity, 2-oxoglutarate metabolic process, valine catabolic process, peroxisome and unfolded protein binding. Pathway significant enrichment analysis indicated 6 signaling pathways that are significant, including steroid biosynthesis, steroid hormone biosynthesis, Parkinson's disease, 2,4-Dichlorobenzoate degradation, and tropane, piperidine and Isoquinoline alkaloid biosynthesis. Among these, eleven selected genes were further verified by qRT-PCR. Our findings provide a comprehensive view on the gene expression profile of M. canis upon berberine treatment, and shed light on its complicated effects on M. canis.
小檗碱是多种药草中的一种天然异喹啉生物碱,对包括犬小孢子菌(M. canis)在内的多种微生物感染具有积极作用。然而,其潜在机制尚不清楚。为了研究氯化小檗碱对犬小孢子菌感染的影响,构建了数字基因表达(DGE)标签谱,并对犬小孢子菌经小檗碱处理后的细胞反应进行了转录组分析。采用Illumina/Hisseq测序技术生成基因表达谱数据,并基于转录组数据进行了以下基因本体(GO)和通路功能富集分析。DGE结果显示,分别从用小檗碱或对照二甲基亚砜(DMSO)孵育的犬小孢子菌中获得了8476945、14256722、7708575、5669955、6565513和9303468个标签。总共映射了8783个基因,两组之间有1890个基因显示出显著变化。与对照组相比,小檗碱处理组中有1030个基因上调,860个基因下调(P<0.05)。此外,通过基因本体功能富集分析确定了23个GO术语,如钙转运ATP酶活性、2-氧代戊二酸代谢过程、缬氨酸分解代谢过程、过氧化物酶体和未折叠蛋白结合。通路显著富集分析表明有6条信号通路显著,包括类固醇生物合成、类固醇激素生物合成、帕金森病、2,4-二氯苯甲酸降解以及托烷、哌啶和异喹啉生物碱生物合成。其中,11个选定基因通过qRT-PCR进一步验证。我们的研究结果提供了犬小孢子菌经小檗碱处理后的基因表达谱的全面视图,并揭示了其对犬小孢子菌的复杂影响。