丁香油联用β-内酰胺类抗生素体外抗耐甲氧西林金黄色葡萄球菌作用研究
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摘要
目的探讨丁香油体外抗耐甲氧西林金黄色葡萄球菌(MRSA)活性,以及与3种β-内酰胺类抗生素(阿莫西林、头孢氨苄和头孢吡肟)联合使用抗MRSA的体外活性。方法采用96孔板微量稀释法分别测定丁香油、丁香油与3种β-内酰胺类抗生素联合使用对2株MRSA标准株和32株MRSA临床分离株的最低抑菌浓度(MIC);微量棋盘法分别测定丁香油与3种β-内酰胺类抗生素的分级抑菌浓度(FIC)指数;生长曲线法分析丁香油以及丁香油和头孢氨苄联合使用对MRSA生长的影响。结果丁香油具有明显的体外抗MRSA活性,MIC为0.25、0.51 mg/m L。丁香油分别与3种β-内酰胺类抗生素(阿莫西林、头孢氨苄和头孢吡肟)联合使用时,FIC指数集中分布在≤0.5和0.5~1,主要表现为协同和相加作用,其中与阿莫西林、头孢氨苄和头孢吡肟的协同率分别为58.8%、97.0%、64.7%;丁香油能明显增强β-内酰胺类抗生素抗MRSA活性,降低β-内酰胺类抗生素的用量,使其MIC50和MIC90降低为单独用药的1/4~1/2和1/2~1。生长曲线结果表明,丁香油能明显延长MRSA延滞期,抑制其生长速度,生长量明显降低,且抑制MRSA生长呈明显量效关系;当丁香油(1/4 MIC)与头孢氨苄(1/4 MIC)联合使用后,MRSA生长几乎被抑制。结论丁香油不仅自身具有较强的抗MRSA活性,还具有协同增效β-内酰胺类抗生素抗MRSA作用,具备治疗MRSA感染新药的开发与应用前景。
Objective To investigate the antimicrobial activities of clove oil alone and in combination with three kinds of β-lactam antibiotics(amoxicillin, cephalexin, and cefepime) against methicillin-resistant Staphylococcus aureus(MRSA). Methods The minimum inhibitory concentrations(MICs) of clove oil alone and in combination with three kinds of β-lactam antibiotics against two standard MRSA and 32 MRSA isolated from clinic were determined by 96-well plate micro-dilution method. The fractional inhibitory concentrations(FICs) of clove oil and three kinds of β-lactam antibiotics were determined by using the micro-chessboard method. The growth curve of MRSA ATCC43300 effected by clove oil or clove oil combined with cephalexin was analyzed. Results Clove oil showed significant anti-MRSA activity in vitro, with MIC of 0.25 mg/m L and 0.51 mg/m L. The FICs were mainly distributed in the range of ≤ 0.5 and 0.5 to 1, and the synergy rates were 58.8%, 97.0%, and 64.7%, when the clove oil was combined with amoxicillin, cephalexin and cefepime, respectively. Clove oil enhanced the activity of β-lactam antibiotics against MRSA significantly, and reduced the dosage of it. The growth curve showed that the clove oil significantly inhibited the growth of MRSA, and prolonged its lag phase. And the effect showed a significant dose-effect relationship. The growth of MRSA was nearly inhibited when 1/4 MIC cefepime combined with 1/4 MIC clove oil. Conclusion Clove oil not only could exhibit strong anti-MRSA activity itself, but could enhance the activity of β-lactam antibiotics against MRSA. Hence, clove oil would be a potential new drug, which can be used as a new drug for preventing and treating MRSA infection.
Objective To investigate the antimicrobial activities of clove oil alone and in combination with three kinds of β-lactam antibiotics(amoxicillin, cephalexin, and cefepime) against methicillin-resistant Staphylococcus aureus(MRSA). Methods The minimum inhibitory concentrations(MICs) of clove oil alone and in combination with three kinds of β-lactam antibiotics against two standard MRSA and 32 MRSA isolated from clinic were determined by 96-well plate micro-dilution method. The fractional inhibitory concentrations(FICs) of clove oil and three kinds of β-lactam antibiotics were determined by using the micro-chessboard method. The growth curve of MRSA ATCC43300 effected by clove oil or clove oil combined with cephalexin was analyzed. Results Clove oil showed significant anti-MRSA activity in vitro, with MIC of 0.25 mg/m L and 0.51 mg/m L. The FICs were mainly distributed in the range of ≤ 0.5 and 0.5 to 1, and the synergy rates were 58.8%, 97.0%, and 64.7%, when the clove oil was combined with amoxicillin, cephalexin and cefepime, respectively. Clove oil enhanced the activity of β-lactam antibiotics against MRSA significantly, and reduced the dosage of it. The growth curve showed that the clove oil significantly inhibited the growth of MRSA, and prolonged its lag phase. And the effect showed a significant dose-effect relationship. The growth of MRSA was nearly inhibited when 1/4 MIC cefepime combined with 1/4 MIC clove oil. Conclusion Clove oil not only could exhibit strong anti-MRSA activity itself, but could enhance the activity of β-lactam antibiotics against MRSA. Hence, clove oil would be a potential new drug, which can be used as a new drug for preventing and treating MRSA infection.
引文
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[12]Chen X,Shang F,Meng Y,et al.Ethanol extract of Sanguisorba officinalis L.inhibits biofilm formation of methicillin resistant Staphylococcus aureus in an ica-dependent manner[J].J Dairy Sci,2015,98(12):8486-8491.
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[15]曾荣,陈金印,林丽超.丁香精油及丁香酚对食品腐败菌的抑菌活性研究[J].江西农业大学学报,2013,35(4):852-857.
[16]Bhuiyan M N I,Begum S,Nandi N C,et al.Constituents of the essential oil from leaves and buds of clove(Syzigium caryophyllatum(L.)Alston)[J].Afr J Plant Sci,2010,6(16):451-454.
[17]马艳玲,李海贤,曾荣.丁香酚对金黄色葡菌球菌抗菌作用的探究[J].中国酿造,2017,36(8):130-133.
[18]马莎莎,赵震宇,刘平怀,等.丁香不同提取方法提取物的抗氧化活性[J].精细化工,2018,35(2):218-227.
[19]Ozcan M M,Arslan D.Antioxidant effect of essential oils of rosemary,clove and cinnamon on hazelnut and poppy oils[J].Food Chem,2011,129(1):171-174.
[20]Kong B,Zhang H,Xiong Y L.Antioxidant activity of spice extracts in a liposome system and in cooked pork patties and the possible mode of action[J].Meat Sci,2010,85(4):772-778.
[21]周建新,许华,金浩.丁香油抑菌效果与抑菌成分的研究[J].食品工业,2003,21(3):24-26.
[22]陈蔚文,李茹柳,叶富强,等.连香胶囊治疗胃院痛的临床观察与药理实验研究[J].中国新药与临床药理,1999,10(5):268-271.
[23]谭延华,何西利,高健,等.丁香酚对氧自由基的清除作用[J].西北药学杂志,1996,11(1):30-31.
[24]朱金段,袁德俊,林新颖.丁香的药理研究现状及临床应用[J].中国药物经济学,2013(1):32-35.
[25]伍振峰,王赛君,杨明,等.中药挥发油提取工艺与装备现状及问题分析[J].中国实验方剂学杂志,2014,20(14):224-227.
[26]朱生墚,马淑影,王小素,等.丁香降气汤治疗反流性食管炎50例临床观察[J].上海中医药杂志,2005,39(1):19-20.
[27]Janardhanan S,Mahendra J,Girija A S,et al.Antimicrobial effects of Garcinia Mangostana on cariogenic microorganisms[J].J Clin Diagn Res,2017,11(1):19-22.
[28]张肖冰,杨事达,姚志成,等.替加环素与5种抗生素对多重耐药鲍曼不动杆菌体外联合药敏实验的研究[J].微生物学杂志,2017,37(5):82-87.
[29]孟佳敏,江汉美,卢金清,等.丁香高效液相指纹图谱研究[J].中国药师,2018,21(10):1762-1765.
[30]徐航,龙娜娜,林琳,等.草果油抗MRSA体外活性研究[J].成都医学院学报,2017,12(3):241-246.
[31]杨滋渊,李巧如,赵院莉,等.丁香挥发油的抗菌作用研究[J].西北药学杂志,2007,22(4):181.
[32]Roemer T,Schneider T,Pinho M G.Auxiliary factors:A chink in the armor of MRSA resistance toβ-lactam antibiotics[J].Curr Opin Microbiol,2013,16(5):538-548.
[2]卢克鹏,宋洪涛.糖肽类抗生素在治疗MRSA感染中的PK/PD与给药方案的研究进展[J].中国医院药学杂志,2014,34(11):952-957.
[3]Klevens R M,Morrison M A,Nadle J,et al.Invasive methicillin-resistant Staphylococcus aureus infections in the United States[J].JAMA,2007,298(15):1763-1771.
[4]de Kraker M E,Wolkewitz M,Davey P G,et al.Clinical impact of antimicrobial resistance in European hospitals:Excess mortality and length of hospital stay related to methicillin-resistant Staphylococcus aureus blood stream infections[J].Antimicrob Agents Chemother,2011,55(4):1598-1605.
[5]Kluytmans J,Struelens M.Meticillin resistant Staphylococcus aureus in the hospital[J].BMJ,2009,338(7693):532-537.
[6]王明强,张思森,刘小军,等.替考拉宁与万古霉素及利奈唑胺治疗MRSA感染患者的疗效评价[J].中华医院感染学杂志,2015,25(17):3923-3924.
[7]Giltner C L,Kelesidis T,Hindler J A,et al.Frequency of susceptibility testing for patients with persistent methicillin-resistant Staphylococcus aureus bacteremia[J].J Clin Microbiol,2014,52(1):357-361.
[8]Stryjewski M E,Corey R G.Methicillin-resistant Staphylococcus aureus:An evolving pathogen[J].Clin Infect Dis,2014,58(S1):10-19.
[9]郝佳,刘哲,张瑜,等.苦参、绿茶及紫苏叶提取物联用对金黄色葡萄球菌的体外活性研究[J].天津中医药大学学报,2014,33(5):296-298.
[10]Gardete S,Tomasz A.Mechanisms of vancomycin resistance in Staphylococcus aureus[J].J Clin Invest,2014,124(7):2836-2840.
[11]Tan X,Yang D,Yang G,et al.The investigation of inhibiting quorum sensing and methicillin-resistant Staphylococcus aureus biofilm formation from Liriodendron hybrid[J].Pak J Pharm Sci,2015,28(3):903-908.
[12]Chen X,Shang F,Meng Y,et al.Ethanol extract of Sanguisorba officinalis L.inhibits biofilm formation of methicillin resistant Staphylococcus aureus in an ica-dependent manner[J].J Dairy Sci,2015,98(12):8486-8491.
[13]Liu I X,Durham D G,Michael R.Baicalin synergy with beta-lactam antibiotics against methicillin-resistant Staphylococcus aureus and other beta-lactam-resistant strains of S.aureus[J].J Pharm Pharmacol,2000,52(3):361-366.
[14]屠寒,江汉美,卢金清,等.丁香药理作用研究进展[J].香料香精化妆品,2015(5):59-62.
[15]曾荣,陈金印,林丽超.丁香精油及丁香酚对食品腐败菌的抑菌活性研究[J].江西农业大学学报,2013,35(4):852-857.
[16]Bhuiyan M N I,Begum S,Nandi N C,et al.Constituents of the essential oil from leaves and buds of clove(Syzigium caryophyllatum(L.)Alston)[J].Afr J Plant Sci,2010,6(16):451-454.
[17]马艳玲,李海贤,曾荣.丁香酚对金黄色葡菌球菌抗菌作用的探究[J].中国酿造,2017,36(8):130-133.
[18]马莎莎,赵震宇,刘平怀,等.丁香不同提取方法提取物的抗氧化活性[J].精细化工,2018,35(2):218-227.
[19]Ozcan M M,Arslan D.Antioxidant effect of essential oils of rosemary,clove and cinnamon on hazelnut and poppy oils[J].Food Chem,2011,129(1):171-174.
[20]Kong B,Zhang H,Xiong Y L.Antioxidant activity of spice extracts in a liposome system and in cooked pork patties and the possible mode of action[J].Meat Sci,2010,85(4):772-778.
[21]周建新,许华,金浩.丁香油抑菌效果与抑菌成分的研究[J].食品工业,2003,21(3):24-26.
[22]陈蔚文,李茹柳,叶富强,等.连香胶囊治疗胃院痛的临床观察与药理实验研究[J].中国新药与临床药理,1999,10(5):268-271.
[23]谭延华,何西利,高健,等.丁香酚对氧自由基的清除作用[J].西北药学杂志,1996,11(1):30-31.
[24]朱金段,袁德俊,林新颖.丁香的药理研究现状及临床应用[J].中国药物经济学,2013(1):32-35.
[25]伍振峰,王赛君,杨明,等.中药挥发油提取工艺与装备现状及问题分析[J].中国实验方剂学杂志,2014,20(14):224-227.
[26]朱生墚,马淑影,王小素,等.丁香降气汤治疗反流性食管炎50例临床观察[J].上海中医药杂志,2005,39(1):19-20.
[27]Janardhanan S,Mahendra J,Girija A S,et al.Antimicrobial effects of Garcinia Mangostana on cariogenic microorganisms[J].J Clin Diagn Res,2017,11(1):19-22.
[28]张肖冰,杨事达,姚志成,等.替加环素与5种抗生素对多重耐药鲍曼不动杆菌体外联合药敏实验的研究[J].微生物学杂志,2017,37(5):82-87.
[29]孟佳敏,江汉美,卢金清,等.丁香高效液相指纹图谱研究[J].中国药师,2018,21(10):1762-1765.
[30]徐航,龙娜娜,林琳,等.草果油抗MRSA体外活性研究[J].成都医学院学报,2017,12(3):241-246.
[31]杨滋渊,李巧如,赵院莉,等.丁香挥发油的抗菌作用研究[J].西北药学杂志,2007,22(4):181.
[32]Roemer T,Schneider T,Pinho M G.Auxiliary factors:A chink in the armor of MRSA resistance toβ-lactam antibiotics[J].Curr Opin Microbiol,2013,16(5):538-548.