没食子酸与头孢类抗生素对胃肠外科术后主要感染病原菌的联合抗菌作用及机理研究  

Study on antibacterial activity and mechanism of Ampelopsis grossedentata flavone extract against pathogenic Vibrio

作　　者：李玲梅 蔡延渠[2,3,4] LI Lingmei;CAI Yanqu(Department of Pharmacy,The Fifth Affiliated Hospital Of Guangzhou Medical University,Guangzhou 510700,Guangdong,China;Center for Drug Research and Development,Guangdong Pharmacological University,Guangzhou 510006,Guangdong,China;Class III Laboratory of Modern Chinese Medicine Preparation,State Administration of Traditional Chinese Medicine of the P.R.C,Guangzhou 510006,Guangdong,China;Key Laboratory of modern Chinese medicine of Education Department of Guangdong Province,Guangzhou 510006,Guangdong,China)

机构地区：[1]广州医科大学附属第五医院药学部,广东广州510700 [2]广东药科大学新药研发中心,广东广州510006 [3]国家中医药管理局中药制剂实验室,广东广州510006 [4]广东省教育厅现代中药重点实验室,广东广州510006

出　　处：《广州医科大学学报》2024年第6期1-10,共10页Academic Journal of Guangzhou Medical University

基　　金：广东省中医药局项目(20242051)。

摘　　要：目的:研究没食子酸联合头孢类抗生素对金黄色葡萄球菌等胃肠外科术后主要感染病原菌的体外抑菌活性和作用机制,为其作为抗菌增效剂进行联合用药提供依据。方法:抗菌活性:采用微量稀释法测定没食子酸(GA)、头孢唑林、头孢呋辛、头孢他啶、头孢曲松、头孢吡肟对金黄色葡萄球菌(SA)、肺炎克雷伯菌(KP)、大肠杆菌(EC)的最低抑菌浓度(MIC)及最低杀菌浓度(MBC),同时采用棋盘稀释法测定不同浓度没食子酸与5种头孢类抗生素的联合抑菌指数(FICI)。作用机制:采用多种方法测定SA、EC的生长曲线、胞外碱性磷酸酶活性、胞外核酸含量、胞外蛋白质浓度、膜电位变化、生物膜清除率、生物膜代谢活性及胞外多糖分泌量等指标,评价没食子酸联合头孢曲松对受试菌株细胞壁、细胞膜及生物膜的影响。结果:GA对SA、KP、EC的MIC、MBC为2.5~5.0、2.5~10.0 mg/mL,5种头孢类抗生素的MIC为0.24~3.91、0.98~7.81、0.98~15.63μg/mL,MBC为0.98~15.63、1.95~31.25、1.95~62.50μg/mL。对SA、KP、EC的FICI依次为1.0~1.125、1.125~1.125、0.625~0.625。SA、EC在GA/CTRX在MIC联浓度下作用6~12 h,菌体生长受到显著抑制,胞外AKP活性显著上升至5.58~7.86、8.19~11.29 U/L,胞外核酸A_(260)显著上升至4.36~5.38、5.22~6.73,胞外蛋白质浓度显著上升至7.18~18.79、14.18~35.07mg/L,膜电位显著下降至185.90~65.07、134.60~80.20 AU,生物膜清除率为30.92%、36.21%,生物膜代谢活性抑制率为35.67%、45.09%,胞外多糖分泌量显著下降至60.54、53.86 mg/L。结论:GA与5种头孢类抗生素联用均可降低抗生素用量,对SA、EC表现出相加效应,其作用机制与破坏菌体细胞壁、细胞膜完整性以及影响生物膜形成有关,可作为新的抗菌增效剂与头孢类抗生素进行联合用药,为临床术后感染性疾病提供新的有效治疗策略。Objective:To study the in vitro antibacterial activity and mechanism of gallic acid combined with cephalosporins against Staphylococcus aureus and other major infectious pathogens after gastrointestinal surgery,so as to provide a basis for its combined use as an antibacterial synergist.Methods:Gallic acid(GA),cefazolin,cefuroxime,ceftazidime,ceftriaxone,cefepime were determined by microdilution method against the minimum inhibitory concentration(MIC)and minimum bactericidal concentration(MBC)of Staphylococcus aureus(SA),Klebsiella pneumoniae(KP)and Escherichia coli(EC).At the same time,the combined inhibitory index(Fici)of gallic acid in different concentrations and five cephalosporin antibiotics was determined.The indexes of growth curve,extracellular alkaline phosphatase activity,extracellular nucleic acid content,extracellular protein concentration,membrane potential change,biofilm clearance,biofilm metabolic activity and extracellular polysaccharide secretion of Staphylococcus aureus and Escherichia coli were determined by various ways,to evaluate the effect of gallic acid combined with ceftriaxone on the cell wall,cell membrane and biofilm of the tested strains.Results:The MIC and MBC of Gallic acid to Staphylococcus aureus,Klebsiella pneumoniae and Escherichia coli are 2.5-5.0 mg/mL and 2.5-10.0 mg/mL,and the MIC of five cephalosporins were 0.24-3.91μg/mL,0.98-7.81μg/mL,0.98~15.63μg/mL,and the MBC were 0.98-15.63μg/mL,1.95-31.25μg/mL,1.95-62.50μg/mL.The Fici of Staphylococcus aureus,Klebsiella pneumoniae and Escherichia coli are 1.0-1.125,1.125-1.125 and 0.625-0.625 respectively.Staphylococcus aureus and Escherichia coli were significantly inhibited 6 to 12 hours by GA/CTRX at the concentration of MIC,and the extracellular AKP activity increased significantly to 5.58-7.86 and 8.19-11.29 U/L,the extracellular nucleic acid A_(260) increased significantly to 4.36-5.38 and 5.22-6.73,the extracellular protein concentration increased significantly to 7.18-18.79 mg/L and 14.18-35.07 mg/L,the membrane potenti

关 键 词：术后感染病原菌 没食子酸 头孢类抗生素 联合抗菌 作用机制 

分 类 号：R965[医药卫生—药理学]