黄芩水煎剂对尿道致病性大肠埃希菌的转录组影响分析  

Effect analysis on water decoction of Scutellaria baicalensis to transcription of uropathogenic Escherichia coli

作　　者：朱健铭 翁幸鐾[2] 姜如金[1] 吴晋兰[3] 贺子龙[4] 姚娜[1] 

机构地区：[1]杭州市余杭区中医院,浙江杭州311106 [2]宁波市第一医院,浙江宁波315010 [3]杭州市余杭区第五人民医院,浙江杭州311100 [4]中国科学院北京基因组研究所基因组科学与信息重点实验室,北京100101

出　　处：《中草药》2017年第9期1791-1801,共11页Chinese Traditional and Herbal Drugs

基　　金：杭州市科技计划引导项目(2012028);杭州市卫生科技计划项目(2012B034);杭州市余杭区重大科技项目(2012006)

摘　　要：目的探讨黄芩水煎剂抑制尿道致病性大肠埃希菌的分子机制。方法采用RNA-seq技术分析黄芩水煎剂对尿道致病性大肠埃希菌临床分离株(NB8)转录组的影响。设黄芩组和阴性对照组2个组,黄芩组用10倍MIC浓度的黄芩水煎剂(62.5 mg/mL)作用于尿道致病性大肠埃希菌NB8株30 min,对照组给等量生理盐水,提取细菌总RNA,去除rRNA,反转录合成cDNA,在HiSeq2000测序平台上进行转录组测序,利用BIGpre、Tophat、Cufflinks等生物信息学工具软件进行转录组数据处理;并将得到的表达谱作差异表达、GO和COG功能富集以及KEGG代谢通路分析。结果黄芩组和对照组之间差异表达基因共有1 665个,其中上调基因为1 169个,下调基因为496个;在黄芩水煎剂的作用下,尿道致病性大肠埃希菌NB8株糖酵解、三羧酸循环和脂肪酸生物合成等关键代谢途径的编码基因以及核糖体蛋白的编码基因显著下调,而细菌趋化性和鞭毛组装途径以及ABC转运蛋白通路相关基因表达显著上调。结论阐明了黄芩水煎剂抑制尿道致病性大肠埃希菌的分子机制,黄芩作用的靶位是糖酵解、三羧酸循环、脂肪酸的生物合成途径和蛋白质的翻译;此外,细菌趋化性、鞭毛组装途径和ABC转运蛋白在细菌对黄芩的应激反应中也具有重要作用。Objective To investigate the molecular mechanism of inhibitory effect of Scutellaria baicalensis to uropathogenic Escherichia coli（UPEC）. Methods RNA-seq was used to analyze transcriptome of UPEC NB8 effected by water decoction of S. baicalensis. S. baicalensis group and negative control group were set up. In S. baicalensis group, UPEC NB8 was disposed by water decoction of 10 times of MIC of S. baicalensis（62.5 mg/mL） for 30 min. While in negative control group, UPEC NB8 was disposed by same amount of normal saline. Then, total RNA of UPEC NB8 was extracted, while rRNA was removed, cDNA was reverse transcriptional synthesized. Transcriptome sequencing was sequenced on HiSeq2000, transcriptome data were analyzed by BIGpre, Tophat, and Cufflinks. Expression profile was analyzed for differential expression, GO, COG functional enrichment analysis, and KEGG metabolic pathway analysis. Results A total of 1665 differentially expressed genes existed between S. baicalensis group and negative control group. Among them, 1169 genes were up-regulated genes, while 496 genes were down-regulated genes. Under the effect of water decoction of S. baicalensis, genes of NB8 down-regulated in glycolysis, Krebs cycle, biosynthesis of fatty acids, and ribosomal proteins, while genes of NB8 up-regulated chemotaxis and assembly path of flagella, and transportation protein ABC pathway. Conclusion The molecular mechanism of the inhibitory effect of S. baicalensis to UPEC is explained. Targets of S. baicalensis to UPEC are glycolysis, Kreb＇s cycle, biosynthesis of fatty acids, and translation of proteins. Besides, chemotaxis, the assembly path of flagella, and transportation protein ABC also play a key role in alarm reaction of S. baicalensis.

关 键 词：黄芩 尿道致病性大肠埃希菌 转录组 RNA-SEQ 差异表达基因 分子机制 

分 类 号：R285.5[医药卫生—中药学]