水飞蓟宾及其同分异构体的抑菌谱及其与抗生素联合抑菌效应探讨
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摘要
目的探讨水飞蓟宾及同分异构体的体外抗菌谱及水飞蓟宾与临床常用抗生素的联合抑菌效应。方法用微量肉汤稀释法测定水飞蓟宾及同分异构体对临床感染常见细菌的6种标准菌株和6种临床分离菌株(74株)的最低抑菌浓度(MIC)。用平板菌落计数法测定不同质量浓度水飞蓟宾对6种标准菌株的生长抑制曲线。用棋盘微量肉汤稀释法进行水飞蓟宾与临床常用抗生素的联合药敏试验,计算联合抑菌指数(FIC),判定水飞蓟宾与抗生素的联合抑菌效应。结果水飞蓟宾对表皮葡萄球菌、金黄色葡萄球菌、粪肠球菌和屎肠球菌标准菌株的MIC为50~400μg/m L,对大肠埃希菌和铜绿假单胞菌标准菌株的MIC均>400μg/m L;对表皮葡萄葡萄球菌临床分离菌株的MIC分别为100、200、400、>400μg/m L,对金黄色葡萄球菌、粪肠球菌和屎肠球菌临床分离菌株的MIC分别为400、>400μg/m L;对大肠埃希菌和铜绿假单胞菌临床分离菌株的MIC均>400μg/m L。水飞蓟宾其他同分异构体对6种标准菌株的MIC为≥400μg/m L。水飞蓟宾对表皮葡萄球菌、金黄色葡萄球菌、粪肠球菌和屎肠球菌标准菌株的生长曲线都有明显抑制作用,抑制效果随药物质量浓度增加而增加,对大肠埃希菌和铜绿假单胞菌标准菌株的生长曲线无影响。水飞蓟宾与青霉素/红霉素联用对革兰阳性试验菌的FIC以0.52或1 Objective To investigate the in vitro antibacterial spectrum of silibinin and its isomers and the combined antibacterial effect of silibinin and commonly-used clinical antibiotics. Methods Micro-broth dilution method was used to determine the minimum inhibitory concentration(MIC) of silibinin and its isomers against six standard strains and six clinical isolates(74 strains) of common bacteria infections. Plate count method was used to measure the growth inhibition curves of different concentrations of silibinin against six standard strains. Checkerboard micro-broth dilution method was used to carry out the combined susceptibility test of silibinin and clinical antibiotics. The fractional inhibitory concentration(FIC) was calculated to determine the combined antibacterial effect of silibinin and antibiotics. Results The MICs of silibinin against standard strains of Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis, and Enterococcus faecium were between 50 μg/m L and 400 μg/m L, and those of silibinin for standard strains of Escherichia coli and Pseudomonas aeruginosa were both greater than 400 μg/m L. The MICs of silibinin for clinical strains, Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis, and Enterococcus faecium were all within the range of 100 μg/m L to 400 μg/m L, and MICs of silibinin against clinical strains of Escherichia coli and Pseudomonas aeruginosa were greater than 400 μg/m L. The MICs of other isomers of silibinin against six standard strains were greater than or equal to 400 μg/m L. Silibinin had a significant inhibitory effect on the growth curve of standard strains of Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus faecalis, and Enterococcus faecium, which increased with increasing drug concentration, and it had no effect on that of Escherichia coli and Pseudomonas aeruginosa. The FIC of silibinin combined with penicillin/erythromycin for gram-positive test bacteria was in the range of 0.5 < FIC ≤ 1 and 1 < FIC ≤ 2, and FIC of silibinin combined with ciprofloxacin or gentamicin for gram-negative test bacteria was in the range of FIC > 2 or 1 < FIC ≤ 2. Conclusion Silibinin has a notable antibacterial activity to gram-positive bacteria with the best inhibitory effect on Staphylococcus epidermidis. The antibacterial activities of silibinin is higher than those of other isomers. The combined antibacterial effect of silibinin and penicillin/erythromycin is mainly additive and irrelevant, while that of silibinin and ciprofloxacin or gentamicin is mainly antagonistic or irrelevant.
引文
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[18]曾伟民,陶超,范晓旭,等.水飞蓟秸秆中水飞蓟素成分分析[J].黑龙江畜牧兽医,2014,7(13):168-170.
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[21]袁琳慧,王迪,谢鲲鹏,等.穿心莲和水飞蓟抑制MRSA41577外排系统的作用机制[J].中国生化药物杂志,2016,36(11):27-29.
[22]金戈,王俊平,王春红,等.水飞蓟宾减轻庆大霉素肾毒性作用的研究[J].中药药理与临床,2013,29(2):51-53.
[2]冯晶晶,王小万,崔月颖.欧洲促进新抗菌药物研发的措施及经验[J].中国药房,2014,25(1):7-9.
[3]李耘,吕媛,薛峰,等.卫生部全国细菌耐药监测网(Mohnarin)2011—2012年革兰阳性菌耐药监测报告[J].中国临床药理学杂志,2014,30(3):251-259.
[4]李耘,吕媛,薛峰,等.卫生部全国细菌耐药监测网(Mohnarin)2011—2012年革兰阴性菌耐药监测报告[J].中国临床药理学杂志,2014,30(3):260-277.
[5]王启,季萍,徐修礼,等.2016年中国14家教学医院革兰阴性杆菌耐药监测分析[J].中华检验医学杂志,2017,40(8):614-622.
[6]靳蕊,徐敏纹,刘莹,等.黄酮类化合物的抑菌作用及其机制的研究[J].继续医学教育,2016,30(8):152-154.
[7]Alvarez-Ordonez A,Carvajal A,Arguello H,et al.Antibacterial activity and mode of action of a commercial citrus fruit extract[J].J Appl Microbiol,2013,115(1):50-60.
[8]李娟,张学顺,傅春升,等.中药抗菌作用的研究进展[J].中国药业,2014,23(2):90-93.
[9]刘一杰,薛永常.植物黄酮类化合物的研究进展[J].中国生物工程杂志,2016,36(9):81-86.
[10]李亚娜,陶庆春.中药抑菌的研究现状及思考[J].国际检验医学杂志,2014,35(2):198-200.
[11]游庭活,刘凡,温露,等.黄酮类化合物抑菌作用研究进展[J].中国中药杂志,2013,38(21):3645-3650.
[12]杜文静,谢亚斌,葛雅琨,等.水飞蓟宾对金黄色葡萄球菌和大肠杆菌的体外抑菌作用的研究[J].吉林化工学院学报,2016,33(7):66-69.
[13]Shukla S K,Dasgupta A,Mehla K,et al.Silibininmediated metabolic reprogramming attenuates pancreatic cancer-induced cachexia and tumor growth[J].Oncotarget,2015,6(38):41146-41161.
[14]Bahmani M,Shirzad H,Rafieian S,et al.Silybum marianum:Beyond hepatoprotection[J].J Evid BasedCompl Altern Med,2015,20(4):292-301.
[15]Wang Z,Zhang L,Wang J,et al.Synergistic interaction of PMAP-36 and PRW4 with aminoglycoside antibiotics and their antibacterial mechanism[J].World J MicrobiolBiotechnol,2014,30(12):3121-3128.
[16]彭齐,谢丽玲,谢俊.中药抑菌机制的研究方法[J].生物技术进展,2015,5(1):66-69.
[17]Lee D G,Kim H K,Park Y,et al.Gram-positive bacteria specific properties of silybin derived from Silybum marianum[J].Arch Pharm Res,2003,26(8):597-600.
[18]曾伟民,陶超,范晓旭,等.水飞蓟秸秆中水飞蓟素成分分析[J].黑龙江畜牧兽医,2014,7(13):168-170.
[19]Evren E,Yurtcu E.In vitro effects on biofilm viability and antibacterial and antiadherent activities of silymarin[J].Folia Microbiol,2015,60(4):351-356.
[20]Jung H J,Lee D G.Synergistic antibacterial effect between silybin and N,N'-dicyclohexylcarbodiimide in clinical Pseudomonas aeruginosa isolates[J].JMicrobiol,2008,46(4):462-467.
[21]袁琳慧,王迪,谢鲲鹏,等.穿心莲和水飞蓟抑制MRSA41577外排系统的作用机制[J].中国生化药物杂志,2016,36(11):27-29.
[22]金戈,王俊平,王春红,等.水飞蓟宾减轻庆大霉素肾毒性作用的研究[J].中药药理与临床,2013,29(2):51-53.