丁香酚对多重耐药大肠杆菌的抑菌活性及其作用机制研究
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摘要
为研究丁香酚对多重耐药大肠杆菌(Multidrug-resistant Escherichia coli,MDR E.coli)的抑菌活性,利用微量二倍稀释法和琼脂扩散法确定丁香酚对多重耐药大肠杆菌的最低抑菌浓度(Minimum inhibitory concentration,MIC)和最低杀菌浓度(Minimum bactericidal concentration,MBC),并利用紫外分光光度法、考马斯亮蓝法等方法初步分析其作用机制。结果表明,丁香酚对MDR E.coli的MIC和MBC分别为2.50 mg/mL和5.00 mg/mL;丁香酚可延长MDR E.coli迟缓期进入对数生长期的进程,改变菌体结构,增加菌株可溶性蛋白的含量和胞外核酸的量,增大菌体培养液中的电导率,对MDR E.coli具有较强的抑菌作用,这些作用可能归因于丁香酚对其细胞结构的破坏。本研究为临床有效缓解或解决多重耐药大肠杆菌的耐药、感染和致死率等问题提供新的思路、新途径,并为其在医药和食品开发领域的应用奠定理论基础。
In order to study the antibacterial activity of eugenol on multidrug-resistant Escherichia coli(MDR E.coli),the minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) of eugenol against MDR E.coli were determined by micro-diluted dilution method and agar diffusion method.UV spectrophotometry and the Coomassie blue method were used to analyze the antibacterial activity.The results showed that the MIC and MBC of eugenol for MDR E.coli were 2.50 mg/mL and 5.00 mg/mL,respectively.Eugenol can prolong the progression of MDR E.coli into the logarithmic growth phase,change the structure of the bacteria and increase the soluble protein content and the amount of extracellular nucleic acids of the strain.Eugenol also increased the conductivity in the culture medium.These may be attributed to the destruction effect of eugenol on cellular structure of the bacteria.The study provides a new way to overcome drug resistance,infection and mortality of MDR E.coli,and lay a theoretical foundation for the application of eugenol in medicine and food development.
In order to study the antibacterial activity of eugenol on multidrug-resistant Escherichia coli(MDR E.coli),the minimum inhibitory concentration(MIC) and minimum bactericidal concentration(MBC) of eugenol against MDR E.coli were determined by micro-diluted dilution method and agar diffusion method.UV spectrophotometry and the Coomassie blue method were used to analyze the antibacterial activity.The results showed that the MIC and MBC of eugenol for MDR E.coli were 2.50 mg/mL and 5.00 mg/mL,respectively.Eugenol can prolong the progression of MDR E.coli into the logarithmic growth phase,change the structure of the bacteria and increase the soluble protein content and the amount of extracellular nucleic acids of the strain.Eugenol also increased the conductivity in the culture medium.These may be attributed to the destruction effect of eugenol on cellular structure of the bacteria.The study provides a new way to overcome drug resistance,infection and mortality of MDR E.coli,and lay a theoretical foundation for the application of eugenol in medicine and food development.
引文
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[2]墙建.中药干预多重耐药菌(MDRO)机制及抗菌谱研究进展[J].医学理论与实践,2018,31(13):1918-1920QIANG Jian.The progress of traditional chinese medicine intervention in multidrug-resistant bacteria(MDRO)mechanism and antibacterial spectrum[J].Medical Theory and Practice,2018,31(13):1918-1920
[3]兰仕梅,何彦蓉,周江南,等.连翘联合抗菌药物对多重耐药大肠杆菌B3的抗菌试验[J].上海畜牧兽医通讯,2018,4:18-21LAN Shi-mei,HE Yan-rong,ZHOU Jiang-nan,et al.Antibacterial test of multidrug-resistant Escherichia coli B3by combined anti-bacterial drugs of Forsythia suspensa[J].Shanghai Journal of Animal Husbandry and Veterinary Medicine,2018,4:18-21
[4]郭永刚,胡立磊,樊克锋,等.大蒜提取液对大肠杆菌和金黄色葡萄球菌的抑菌效果观察[J].中兽医学杂志,2018,3:14-15GUO Yong-gang,HU Li-lei,FAN Ke-feng,et al.Antibacterial effect of garlic extract on Escherichia coli and Staphylococcus aureus[J].Chinese Journal of Veterinary Medicine,2018,3:14-15
[5]吕世明,谭艾娟,曹敏,等.7种天然化合物对产ESBLs大肠杆菌的增敏机制研究[J].畜牧与兽医,2016,48(4):35-38LU Shi-ming,TAN Ai-juan,CAO Min,et al.Study on the sensitizing mechanism of seven natural compounds to Escherichia coli producing ESBLs[J].Animal Husbandry and Veterinary Medicine,2016,48(4):35-38
[6]国家药典委员会.中华人民共和国药典一部[M].北京:中国医药科技出版社,2015:4National Pharmacopoeia Commission.Pharmacopoeia of the People's Republic of China[M].Beijing:China Medical Science and Technology Press,2015:4
[7]赵晨曦,梁逸曾,李晓宁.丁香挥发油化学成分与抗菌活性研究(英文)[J].天然产物研究与开发,2006,18(3):381-385ZHAO Chen-zhen,LIANG Yi-zeng,LI Xiao-ning.Study on chemical constituents and antibacterial activity of volatile oil from Clove[J].Natural Product Research and Development,2006,18(3):381-385
[8]李巧如,刘宗智,任健康,等.丁香提取物抗菌机理的探讨[J].西北药学杂志,2001,16(6):261-263LI Qiao-ru,LIU Zong-zhi,REN Jian-kang,et al.Discussion on antibacterial mechanism of clove extract[J].Northwest Pharmaceutical Journal,2001,16(6):261-263
[9]孔晓军,刘希望,李剑勇,等.丁香酚的药理学作用研究进展[J].湖北农业科学,2013,52(3):508-511KONG Xiao-jun,LIC Xi-wang,LI Jian-yong,et al.Advance in pharmacological research of eugenol[J].Hubei Agricultural Sciences,2013,52(3):508-511
[10]沈勇根,蔡志鹏,江慎华,等.丁香非挥发性成分抗氧化活性[J].农业机械学报,2012,43(10):131-137SHEN Yong-gen,CAI Zhi-peng,JIANG Shen-hua,et al.Antioxidant activity of non-volatile components of Clove[J].Transactions of the Chinese Society of Agricultural Machinery,2012,43(10):131-137
[11]刘欣,徐娟,洪纯,等.丁香酚和大蒜素分别与亚胺培南联用对多耐药鲍曼不动杆菌的体外抑菌实验研究[J].抗感染药学,2017,14(1):17-20LIU Xin,XU Juan,HONG Chun,et al.Experimental Study on Bacteriostasis of Multidrug Resistant Acinetobacter baumannii in vitro combined by eugenol and allicin with tienam[J].Anti Infect Pharm,2017,14(1):17-20
[12]Prasanth R,Viswanathan P.Anti-virulence potential of eugenol-rich fraction of Syzygium aromaticum against multidrug resistant uropathogens isolated from catheterized patients[J].Avicenna Journal of Phytomedicine,2018:1-11
[13]Das B,Mandal D,Dash S K,et al.Eugenol provokes ROS-mediated membrane damage-associated antibacterial activity against clinically isolated multidrug-resistant Staphylococcus aureus strains[J].Infectious Diseases:Research and Treatment,2016,9
[14]王莹莹,付小宝,于聪,等.6味中草药及配伍方剂提取物对3株大肠杆菌的体外抑菌试验[J].饲料博览,2013,4:43-45WANG Ying-ying,FU Xiao-bao,YU Cong,et al.Test of in vitro antibacteria of E.coli to six Chinese medicine and extract of compatibility prescription[J].Feed Expo,2013,4:43-45
[15]刘延昌.甜茶护齿含片对口腔4种常居菌的体外实验研究[D].广西:广西医科大学,2012LIU Yan-chang.In vitro study of four kinds of resident bacteria in the oral cavity of sweet tea teeth[D].Guangxi:Guangxi Medical University,2012
[16]C Z Chen,S L Cooper.Interactions between den-drimer biocides and bacterial membrances[J].Biomaterial,2002,23(16):3359-3368
[17]Ying-Qiu Li,Qing Han,Jian-Ling Feng,et al.Antibacterial characteristics and mechanisms of?-polylysine against Escherichia coli and Staphylococcus aureus[J].Food Control,2014,43
[18]张涛,陈凡,盖青青,等.离子液与蛋白质和核酸相互作用的研究[J].化学进展,2011,23(10):2132-2139ZHANG Tao,CHEN Fan,GAI Qing-qing,et al.Ionic liquids and protein and nucleic acid interaction[J].Progress in Chemistry,2011,23(10):2132-2139