复方黄柏液涂剂抑制铜绿假单胞菌毒力因子表达的作用机制  

Study on the inhibitory effect of Compound Cortex Phellodendri Liquid on the expression of virulence factors of Pseudomonas aeruginosa

作　　者：杨伟峰[1] 李金泽 崔开宇 李东影[1] 李露 王毅[1] YANG Wei-feng;LI Jin-ze;CUI Kai-yu(Experimental Research Center,China Academy of Chinese Medical Sciences,Beijing(100700),China)

机构地区：[1]中国中医科学院医学实验中心,北京100700 [2]吉林农业大学中药材学院,长春130118

出　　处：《中国中西医结合外科杂志》2023年第3期299-306,共8页Chinese Journal of Surgery of Integrated Traditional and Western Medicine

基　　金：“中医药现代化研究”重点专项(2019YFC1709305);中国中医科学院科技创新工程中西医结合基础重大攻关项目(CI2021A00601);中国中医科学院医学实验中心基本科研业务项目(zz2018018);李连达院士工作站专项经费项目(HX2017001)。

摘　　要：目的:基于网络药理学及体外实验探讨复方黄柏液涂剂抑制铜绿假单胞菌(PAO1)毒力因子表达的作用机制。方法:通过文献检索收集复方黄柏液的有效化学成分,利用STITCH和Pharmmapper数据库进行预测,获得药物抑制PAO1毒力相关的潜在作用靶点,Cytoscape3.7.2构建“中药-成分-靶点”网络;联合STRING数据库构建蛋白质相互作用(PPI)网络图并筛选出核心靶点;利用DAVID对潜在作用靶点进行生物功能与通路富集分析;基于网络分析结果,选择PAO1主要致病毒力因子(碱性蛋白酶、绿脓菌素、细菌生物膜)进行检测,验证复方黄柏液对PAO1毒力因子的影响。结果:复方黄柏液中共筛选出62个抗PAO1的有效成分,潜在作用靶点31个,其中Cynaroside、forsythenside A、Lonijaposide E等为复方黄柏液的主要成分。PPI网络分析得到phnW、lysC、aprA、murC、glmS、gbuA、eta、pvdA、adk、ftsZ共10个核心靶点,KEGG通路注释分析显示与D-丙氨酸代谢、氨基糖和核苷酸糖代谢、代谢途径、精氨酸和脯氨酸代谢和万古霉素耐药信号通路有关。体外实验验证表明,1/2~1/8原液浓度的复方黄柏液对PAO1生长均无影响;与空白对照组比较,1/4~1/8原液浓度复方黄柏液可有效抑制碱性蛋白酶,差异有统计学意义(P<0.05),1/4~1/16浓度复方黄柏液可有效抑制绿脓菌素的产生和成熟期的细菌生物膜,差异有统计学意义(P<0.01)。结论:复方黄柏液可抑制PAO1多种致病毒力因子,从而治疗临床铜绿假单胞菌所致感染。Objective The aim of this study was to investigate the inhibitory effect of Compound Cortex Phellodendri Liquid on the virulence expression of Pseudomonas aeruginosa(PAO1)using network pharmacology and in vitro experiments.Methods The effective chemical constituents of Compound Cortex Phellodendri Liquid were collected through literature search,and the potential targets related to PAO1 virulence were obtained by using STITCH and Pharmmapper databases for prediction.Chinese material medica-ingredients-targets network was constructed with Cytoscape3.7.2.Combine STRING database,construct PPI network map and screen out core targets.Use DAVID database to carry out biological function and pathway enrichment analysis of potential targets.Based on the results of network analysis,the alkaline protease,pyocyanin,and biofilm index were selected to verify the effect of Compound Cortex Phellodendri Liquid on virulence expression of Pseudomonas aeruginosa.Results A total of 62 active components and 31 potential targets of Compound Cortex Phellodendri Liquid against PAO1 were screened.Chinese material medica-ingredients-targets network diagram shows Cynaroside,ForsyThenside A,LoniJaposide E,etc.may be Compound Cortex Phellodendri Liquid important ingredients.PPI network analysis revealed core targets such as phnW,lysC,aprA,murC,glmS,gbuA,eta,pvdA,adk,ftsZ.The KEGG pathway enrichment analysis showed that mainly exerted effect by regulating the D-Alanine metabolism,Amino sugar and nucleotide sugar metabolism,Metabolic pathways,Arginine and proline metabolism,Vancomycin metabolism related to metabolism.The experiments show that there has no inhibitory effect on PAO1 at 1/2~1/8 original solution of Compound Cortex Phellodendri Liquid,but the 1/4-1/8 original solution of Compound Cortex Phellodendri Liquid can inhibited the expression of viral alkaline proteases(P<0.05),1/4-1/16 original solution of Compound Cortex Phellodendri Liquid can also inhibited the expression of pyocyanin and mature biofilms(P<0.01).Conclusion Compound Cort

关 键 词：网络药理学 复方黄柏液涂剂 铜绿假单胞菌 毒力因子 

分 类 号：R289.6[医药卫生—方剂学]