青蒿素及其衍生物的抗菌活性初步研究
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摘要
为了检测青蒿素及其衍生物青蒿琥酯和双氢青蒿素的抗菌活性,该实验采用平皿打孔法和肉汤稀释法检测青蒿素、青蒿琥酯和双氢青蒿素在不同浓度和不同作用时间(8,16,24,32 h)对革兰阴性菌(大肠杆菌)和革兰阳性菌(金黄色葡萄球菌)的抗菌效果,并分别以链霉素和青霉素为阳性对照药。平皿打孔法检测发现,与5 mmol·L~(-1)青蒿琥酯或双氢青蒿素不同,5 mmol·L~(-1)青蒿素24 h对大肠杆菌无明显抑菌圈出现。肉汤稀释法发现,双氢青蒿素对大肠杆菌的抗菌活性比青蒿琥酯和青蒿素都强,24 h的IC_(50)为155.9μmol·L~(-1),青蒿琥酯为370.0μmol·L~(-1),而青蒿素没有发现抗菌活性。青蒿琥酯和双氢青蒿素的抗菌活性均在16～24 h最强,而青蒿素的抗菌活性则出现在8～16 h。对于金黄色葡萄球菌,双氢青蒿素没有检测到明显的抗菌活性。该实验证明,青蒿素、青蒿琥酯和双氢青蒿素对大肠杆菌的抗菌活性为双氢青蒿素>青蒿琥酯>青蒿素;青蒿素与其衍生物表现出不同的抗菌动力学特征。双氢青蒿素对金黄色葡萄球菌没有明显抗菌活性。
In this study,in order to detect the antimicrobial activity of artemisinin and its derivatives artesunate and dihydroartemisinin,two methods including broth dilution and plate punching method were used to detect the antibacterial activity against gram-negative bacteria(Escherichia coli)and gram-positive bacteria(Staphylococcus aureus)of artemisinin,dihydroartemisinin and artesunate at various concentrations within 5 mmol·L~(-1)and at four time points(8,16,24,32 h).Two antibacterial positive drugs,streptomycin against E.coli and penicillin against S.aureus,were used as positive controls.Plate punching method showed that,unlike the results of 5 mmol·L~(-1)dihydroartemisinin or artesunate,no inhibition zone was detected at the same concentration of artemisinin after 24 h-treatment against E.coli.Broth dilution method showed that,the antibacterial activity of dihydroartemisinin against E.coli.was stronger than those of both artesunate and artemisinin;IC_(50)at24 h-treatment was 155.9μmol·L~(-1)for dihydroartemisinin,370.0μmol·L~(-1)for artesunate and none for artemisinin.Interestingly,dihydroartemisinin and artesunate showed the strongest antibacterial activity between 16-24 h,while artemisinin showed relatively stronger antibacterial activity between 8-16 h.Dihydroartermisinin showed no antibacterial activity against S.aureus.Above all,the antibacterial activity of artemisinins against E.coli is dihydroartemisinin>artesunate>artemisinin.Artemisinin and its derivatives have showed different antibacterial kinetics,and no antibacterial activity against S.aureus.has been detected with dihydroartemisinin.
In this study,in order to detect the antimicrobial activity of artemisinin and its derivatives artesunate and dihydroartemisinin,two methods including broth dilution and plate punching method were used to detect the antibacterial activity against gram-negative bacteria(Escherichia coli)and gram-positive bacteria(Staphylococcus aureus)of artemisinin,dihydroartemisinin and artesunate at various concentrations within 5 mmol·L~(-1)and at four time points(8,16,24,32 h).Two antibacterial positive drugs,streptomycin against E.coli and penicillin against S.aureus,were used as positive controls.Plate punching method showed that,unlike the results of 5 mmol·L~(-1)dihydroartemisinin or artesunate,no inhibition zone was detected at the same concentration of artemisinin after 24 h-treatment against E.coli.Broth dilution method showed that,the antibacterial activity of dihydroartemisinin against E.coli.was stronger than those of both artesunate and artemisinin;IC_(50)at24 h-treatment was 155.9μmol·L~(-1)for dihydroartemisinin,370.0μmol·L~(-1)for artesunate and none for artemisinin.Interestingly,dihydroartemisinin and artesunate showed the strongest antibacterial activity between 16-24 h,while artemisinin showed relatively stronger antibacterial activity between 8-16 h.Dihydroartermisinin showed no antibacterial activity against S.aureus.Above all,the antibacterial activity of artemisinins against E.coli is dihydroartemisinin>artesunate>artemisinin.Artemisinin and its derivatives have showed different antibacterial kinetics,and no antibacterial activity against S.aureus.has been detected with dihydroartemisinin.
引文
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[14]Zhou F W,Lei H S,Li F,et al. Design,synthesis,and biological evaluation of dihydroartemisinin-fluoroquinolone conjugates as a novel type of potential antitubercular agents[J]. Bloogr Med Chem Lett,2014,24(8):1912.
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[16]Li B,Yao Q,Pan X C,et al. Artesunate enhances the antibacterial effect of beta-lactam antibiotics against Escherichia coli by increasing antibiotic accumulation via inhibition of the multidrug efflux pump system Acr AB-TolC[J]. J Antimicrobial Chemother,2011,66(4):769.
[17]丁艳娇,左国营,郝小燕. 8种滇产中草药体外抑菌活性筛选[J].贵阳医学院学报,2013,38(2):111.
[18]王春娟,左国营,王根春,等. 26种云南中草药的体外抑菌活性筛选[J].中华中医药杂志,2014,21(1):113.
[19]曾富强,王斌,李旭廷,等. 32种中药水提取物的体外抑菌活性研究[J].四川畜牧兽医,2012,39(11):25
[20]赵凌旭,王蕾.中药黄檗对金黄色葡萄球菌体外抑菌效果的初步研究[J].检验医学,2015,30(9):886.
[21]Esimone C O,Adikwu M U,Nwafor S V,et al. In vitro antimicrobial interactions of arthemeter with some 4-quinolones[J].Boll Chim Farm,2002,141(5):385.
[22]Berman P A,Adams P A. Artemisinin enhances heme-catalysed oxidation of lipid membranes[J]. Free Radic Biol Med,1997,22(7):1283.
[23]Zhang F,Jr G D,Meshnick S R. Hemin-catalyzed decomposition of artemisinin(qinghaosu)[J]. Biochem Pharmacol,1992,43(8):1805.
[24]Chen Y,Zheng J M,Zhu S M,et al. Evidence for hemin inducing the cleavage of peroxide bond of artemisinin(Qinghaosu):cyclic voltammetry and in situ FT IR spectroelectrochemical studies on the reduction mechanism of artemisinin in the presence of hemin[J]. Electrochim Acta,1999,44(14):2345.
[25]Kapetanaki S,Varotsis C. Fourier transform infrared investigation of non-heme Fe(Ⅲ)and Fe(Ⅱ)decomposition of artemisinin and of a simplified trioxane alcohol[J]. J Med Chem,2001,44(19):3150.
[26]Taranto A G,Garnerio J W M,de Olivveira F G,et al. The role of C-centered radicals on the mechanism of action artemisinin[J].J Mol Struct,2002,580:207.
[2]郭媛,李凯,姜晓慧,等.低浓度双氢青蒿素对恶性疟原虫3D7株干预效应观察[J].中国中药杂志,2018,43(16):3397.
[3]Lam N S,Long X,Su X Z,et al. Artemisinin and its derivatives in treating helminthic infections beyond schistosomiasis[J].Pharmacol Res,2018,133:77.
[4]Efferth T. Beyond malaria:the inhibition of viruses by artemisinin-type compounds[J]. Biotechnol Adv,2018,36(6):1730.
[5]Loo C S,Lam N S,Yu D,et al. Artemisinin and its derivatives in treating protozoan infections beyond malaria[J]. Pharmacol Res,2017,117:192.
[6]岑彦艳,赵祎博,李攀,等.青蒿琥酯的药代动力学以及相关药理作用研究进展[J].中国中药杂志,2018,43(19):3970.
[7]赵宇平,王慧,杨光,等.基于文本挖掘技术探索青蒿的药理作用规律[J].中国中药杂志,2016,41(16):3072.
[8]李倩,吴叶宽,黄建国.青蒿素对根瘤菌化感效应研究[J].中国中药杂志,2011,36(24):3428.
[9]Jiang W,Li B,Zheng X,et al. Artesunate in combination with oxacillin protect sepsis model mice challenged with lethal live methicillin-resistant Staphylococcus aureus(MRSA)via its inhibition on proinflammatory cytokines release and enhancement on antibacterial activity of oxacillin[J]. Int Immuno Pharmacol,2011,11(8):1065.
[10]Slade D,Galal A M,Gul W,et al. Antiprotozoal,anticancer and antimicrobial activities of dihydroartemisinin acetal dimers and monomers[J]. Bloorgan Med Chem,2009,17(23):7949.
[11]Yi S,Qin R,Pan X,et al. Design of new antibacterial enhancers based on Acr B's structure and the evaluation of their antibacterial enhancement activity[J]. Int J Mol Sci,2016,17(11):1934.
[12]Wu C,Liu J,Pan X,et al. Design,synthesis and evaluation of the antibacterial enhancement activities of amino dihydroartemisinin derivatives[J]. Molecules,2013,18(6):6866.
[13]文学.青蒿素抗菌作用研究[J].人参研究,2009,21(4):38.
[14]Zhou F W,Lei H S,Li F,et al. Design,synthesis,and biological evaluation of dihydroartemisinin-fluoroquinolone conjugates as a novel type of potential antitubercular agents[J]. Bloogr Med Chem Lett,2014,24(8):1912.
[15]Jiang W,Li B,Zheng X,et al. Artesunate has its enhancement on antibacterial activity ofβ-lactams via increasing the antibiotic accumulation within methicillin-resistant Staphylococcus aureus(MRSA)[J]. J Antibiot,2013,66(6):339.
[16]Li B,Yao Q,Pan X C,et al. Artesunate enhances the antibacterial effect of beta-lactam antibiotics against Escherichia coli by increasing antibiotic accumulation via inhibition of the multidrug efflux pump system Acr AB-TolC[J]. J Antimicrobial Chemother,2011,66(4):769.
[17]丁艳娇,左国营,郝小燕. 8种滇产中草药体外抑菌活性筛选[J].贵阳医学院学报,2013,38(2):111.
[18]王春娟,左国营,王根春,等. 26种云南中草药的体外抑菌活性筛选[J].中华中医药杂志,2014,21(1):113.
[19]曾富强,王斌,李旭廷,等. 32种中药水提取物的体外抑菌活性研究[J].四川畜牧兽医,2012,39(11):25
[20]赵凌旭,王蕾.中药黄檗对金黄色葡萄球菌体外抑菌效果的初步研究[J].检验医学,2015,30(9):886.
[21]Esimone C O,Adikwu M U,Nwafor S V,et al. In vitro antimicrobial interactions of arthemeter with some 4-quinolones[J].Boll Chim Farm,2002,141(5):385.
[22]Berman P A,Adams P A. Artemisinin enhances heme-catalysed oxidation of lipid membranes[J]. Free Radic Biol Med,1997,22(7):1283.
[23]Zhang F,Jr G D,Meshnick S R. Hemin-catalyzed decomposition of artemisinin(qinghaosu)[J]. Biochem Pharmacol,1992,43(8):1805.
[24]Chen Y,Zheng J M,Zhu S M,et al. Evidence for hemin inducing the cleavage of peroxide bond of artemisinin(Qinghaosu):cyclic voltammetry and in situ FT IR spectroelectrochemical studies on the reduction mechanism of artemisinin in the presence of hemin[J]. Electrochim Acta,1999,44(14):2345.
[25]Kapetanaki S,Varotsis C. Fourier transform infrared investigation of non-heme Fe(Ⅲ)and Fe(Ⅱ)decomposition of artemisinin and of a simplified trioxane alcohol[J]. J Med Chem,2001,44(19):3150.
[26]Taranto A G,Garnerio J W M,de Olivveira F G,et al. The role of C-centered radicals on the mechanism of action artemisinin[J].J Mol Struct,2002,580:207.