EGCG对奶牛子宫内膜炎致病菌抗菌作用及网络药理学的研究  

Antibacterial effect of EGCG on bovine endometritis-related bacteria and network pharmacological study

作　　者：张泽辉 吴运谱 崔子寅 梁喜才 王杏 王春田[1] 刘明春[2] ZHANG Ze-hui;WU Yun-pu;CUI Zi-yin;LIANG Xi-cai;WANG Xing;WANG Chun-tian;LIU Ming-chun(College of Laboratory Animal Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;College of Animal Science and Veterinary Medicine,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]辽宁中医药大学实验动物医学院,辽宁沈阳110847 [2]沈阳农业大学动物科学与医学学院,辽宁沈阳110866

出　　处：《中国预防兽医学报》2024年第4期411-417,共7页Chinese Journal of Preventive Veterinary Medicine

基　　金：辽宁省民生科技计划项目(2021JH2/10300032);辽宁中医药大学博士科研启动项目(2100221004、2100223016、2100223015);辽宁中医药大学大学生创新创业训练项目(X202210162017)。

摘　　要：为探究表没食子儿茶素没食子酸酯(EGCG)对细菌性奶牛子宫内膜炎的治疗作用机制,本研究采用微量肉汤稀释法测定EGCG对4种牛子宫内膜炎致病菌(金黄色葡萄球菌、化脓隐秘杆菌、大肠杆菌和铜绿假单胞菌)标准株的最小抑菌浓度(MIC)、最小杀菌浓度(MBC),将各菌液稀释100倍后,采用微量结晶紫法测定EGCG对各菌的最小生物被膜(BF)抑制浓度(MBIC);基于网络药理学分析并筛选EGCG治疗奶牛子宫内膜炎的核心靶点,并进行基因本体(GO)和京都基因与基因组百科全书(KEGG)信号通路的富集分析,采用AutoDock Tool软件进行EGCG与核心靶点的分子对接验证。结果显示:EGCG对4种子宫内膜炎相关致病菌均有一定的抗菌活性,其中对革兰氏阳性菌的抗菌作用较强;EGCG在1 MIC~2 MIC的浓度范围内均能有效抑制4株标准株BF的形成。通过网络药理学筛选出肿瘤坏死因子(TNF)和基质金属蛋白酶9 (MMP9)等为EGCG治疗子宫内膜炎的核心靶点;GO分析结果显示含胶原的细胞外基质通路可能是EGCG治疗子宫内膜炎的主要通路之一;KEGG富集分析显示转化生长因子-β (TGF-β)、分泌抵抗、雌激素等信号通路在EGCG治疗奶牛子宫内膜炎的机制中发挥关键作用;分子对接验证试验结果显示EGCG可与上述核心靶点稳定结合。上述结果表明EGCG对4种奶牛子宫内膜炎致病菌表现出抗菌活性和抑制其BF形成的作用,初步揭示了EGCG治疗奶牛子宫内膜炎的可能作用靶点及其作用机制,本研究为EGCG在奶牛生产中的临床应用提供了科学依据。This study was conducted to investigate the mechanism of the therapeutic effect of epigallocatechin gallate(EGCG)on bacterial endometritis in dairy cows.We determined the minimum inhibitory concentration(MIC),minimum bactericidal concentration(MBC),and minimum biofilm inhibitory concentration(MBIC)of EGCG against four standard strains of pathogenic bacteria(Staphylococcus aureus,Trueperella pyogenes,Escherichia coli,and Pseudomonas aeruginosa)in vitro.Core targets of EGCG for treating endometritis in dairy cows were screened through network pharmacology.The gene ontology(GO)function and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed for targets of EGCG-endometritis intersection.Finally,AutoDock Tools was used for molecular docking to verify the binding activity of EGCG and core targets.The results suggested that EGCG exhibits a particular antimicrobial activity against the four endometritis-associated pathogens,especially having the most inhibitory effect on Gram-positive bacteria.Moreover,EGCG could effectively inhibit biofilm formation of the four standard strains in the concentration range of 1MIC to 2MIC.Two core targets,tumor necrosis factor(TNF)and matrix metalloprotein 9(MMP9),were screened by network pharmacology.GO analysis showed that collagen-containing extracellular matrix pathway may be one of the major pathways for EGCG treatment of endometritis.KEGG enrichment analysis showed that three signal pathways are vital:transforming growth factor-β(TGF-β),endocrine resistance,and estrogen.Molecular docking results showed that EGCG is stable enough to bind to the two core targets.In this study,EGCG has demonstrated antimicrobial activity and anti-biofilm effect against four endometritis-associated pathogens in dairy cows.Our data preliminarily revealed the possible targets of the effect of EGCG for treating endometritis in dairy cows and its mechanism,which provides a reference for further clinical application of EGCG in dairy cow production.

关 键 词：奶牛子宫内膜炎 致病菌 EGCG 网络药理学 信号通路 

分 类 号：S853.74[农业科学—临床兽医学]