碘醚柳胺增强黏菌素对肺炎克雷伯菌抗菌活性机制的转录组学分析  

Transcriptomic analysis of the mechanism of rafoxanide enhanced antibacterial activity of colistin against Klebsiella pneumoniae

作　　者：韩荣嘉 张俊锴 刘佩仪 苗青青 翟亚军 胡功政[1] 吴华[1] HAN Rongjia;ZHANG Junkai;LIU Peiyi;MIAO Qingqing;ZHAI Yajun;HU Gongzheng;WU Hua(College of Veterinary Medicine,Henan Agricultural University,Zhengzhou 450046,China)

机构地区：[1]河南农业大学动物医学院,郑州450046

出　　处：《黑龙江畜牧兽医》2024年第9期13-17,115-117,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31772800);河南省科技攻关项目(222102110203);河南省高校科技创新团队支持计划项目(23IRTSTHN021)。

摘　　要：为了了解碘醚柳胺(rafoxanide,RAF)增强黏菌素(colistin,COL)对肺炎克雷伯菌抗菌活性的分子机制,试验采用微量肉汤稀释法初步判断RAF和COL单独或联合后对肺炎克雷伯菌KP1的最小抑菌浓度(MIC)及两药联合的部分抑菌浓度(FIC)指数,然后应用转录组学方法对RAF和COL联合作用4 h后的耐药基因聚类热图、差异表达基因火山图、KEGG通路富集、KEGG功能注释及小分子代谢和转运的差异表达基因进行分析。结果表明:COL对KP1的MIC为128μg/mL,与RAF联用后MIC降低至0.25μg/mL。RAF和COL具有协同作用,FIC指数为0.007 8~0.008 8。与单药COL比较,两药联合后耐药基因bla_(CTX-M)、bla_(SHV)、aac(3)-Ⅱd、aph3-Ⅰ、aac(6′)-Ⅰb、gyrA、gyrB和parC的表达量均下降;共下调了2 017个差异表达基因;下调基因中富集程度和基因数目较为显著的通路有氨酰基-tRNA合成、卟啉与叶绿素代谢、脂多糖生物合成、氧化磷酸化和核糖体;显著富集的通路主要有碳水化合物代谢、辅助因子和维生素代谢、膜转运、氨基酸代谢、能量代谢和信号转导;甘油代谢、核糖体转运、氧化磷酸化、三羧酸循环和抗氧化等与小分子代谢和转运有关的差异表达基因下调。说明RAF可能通过影响肺炎克雷伯菌的代谢和相关小分子的转运功能来增强COL的抗菌活性。In order to understand the molecular mechanism of rafoxanide(RAF) enhancing the antibacterial activity of colistin(COL) against Klebsiella pneumoniae,the minimum inhibitory concentration(MIC) and the partial inhibitory concentration(FIC) index of RAF and COL alone or combined against Klebsiella pneumoniae were determined by microbroth dilution method.Then,transcriptomic methods were used to analyze the clustering heat map of drug resistance genes,differential expression gene volcano map,KEGG pathway enrichment,KEGG functional annotation,and differential gene expression of small molecule metabolism and transport after the combination of RAF and COL for 4 h.The results showed that the MIC value of COL against KP1 was 128 μg/mL,which could be reduced to 0.25 μg/mL when combined with RAF.RAF and COL had a synergistic effect with 0.0078 ≤FIC≤0.008 8.Compared with COL alone,the expression of resistant genes bla_(CTX-M),bla_(SHV),aac(3)-Ⅱd,aph3-Ⅰ,aac(6′)-Ⅰb,gyrA,gyrB,and parC decreased after combination therapy.A total of 2 017 differentially expressed genes were downregulated.The pathways of significant gene enrichment and gene number in down-regulated genes were aminoacyl-TRNA synthesis,porphyrin and chlorophyll metabolism,lipopolysaccharide biosynthesis,oxidative phosphorylation and ribosome.The pathways of significant enrichment mainly include carbohydrate metabolism,cofactor and vitamin metabolism,membrane transport,amino acid metabolism,energy metabolism and signal transduction.The differentially expressed genes related to small molecule metabolism and transport,such as glycerol metabolism,ribosome transport,oxidative phosphorylation,tricarboxylic acid cycle and antioxidant,were down-regulated.This indicated that RAF might enhance the antibacterial activity of COL by affecting the metabolism and transport functions of Klebsiella pneumoniae and related small molecules.

关 键 词：碘醚柳胺 黏菌素 肺炎克雷伯菌 转录组学分析 机制 

分 类 号：S852.612[农业科学—基础兽医学] S859.796[农业科学—兽医学]