青蒿琥酯对大肠杆菌临床分离株抗生素敏感性的影响研究
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摘要
探讨青蒿琥酯(Artesunate,ARS)的体外抗菌活性及其与临床常用抗生素联合对临床耐药大肠杆菌(Escherichia coli,E.coli)的作用效果。采用微量肉汤稀释法测定ARS与7种抗生素分别对6株临床分离E.coli的敏感性;采用棋盘稀释法测定ARS分别与7种抗生素联合对耐药E.coli的分级抑菌浓度(fractional inhibitory concentration,FIC),判定联合抑菌效应;采用平皿打孔法测定ARS预处理对抗生素抑菌圈直径的影响。结果显示,单独使用ARS无明显抑菌作用,但当ARS≥512μg/m L时,与抗生素联用可显著降低抗生素MIC值,其中除链霉素FIC指数为0.6,表现为相加作用外,其余抗菌药FIC指数均小于0.5,表现为协同效应。使用512μg/m L和1024μg/m L ARS前处理耐药E.coli仅增加链霉素和黏菌素的抑菌圈直径。结果表明,ARS本身不具有抑菌活性,但与受测试抗生素同时使用可显著增加抗生素对耐药E.coli的抗菌效果,而ARS预处理对受测试抗生素的增敏效果有限。
The antibacterial activity of artesunate( ARS) in vitro and its effect on clinical drug-resistant Escherichia coli( E.coli) in combination with antibiotics were investigated. The sensitivity of ARS and 7 antibiotics to 6 clinical isolates of E. coli were determined by micro-broth dilution method. The fractional inhibitory concentration( FIC) of ARS combining with 7 antibiotics against resistant E.coli was determined by checkerboard dilution method. The plate perforation method was used to study the effect of ARS pretreatment on the diameter of antibiotic inhibition zone. ARS had no obvious antibacterial effect when used alone. However,when combined with antibiotics,ARS( ≥512 μg/m L) could significantly reduce the MIC value of antibiotics. The FIC indexes of6 antibiotics were all less than 0.5 except for streptomycin,showing synergistic effects. In addition,pretreatment by 512 μg/m L and 1024 μg/m L ARS increased the inhibition zone diameter of both streptomycin and colistin against resistant E. coli, with no effects to other tested antibiotics. Taken together, ARS has no inherent antibacterial activity,but it can significantly enhance the antibacterial effect of antibiotics against drug-resistant E.coli.
The antibacterial activity of artesunate( ARS) in vitro and its effect on clinical drug-resistant Escherichia coli( E.coli) in combination with antibiotics were investigated. The sensitivity of ARS and 7 antibiotics to 6 clinical isolates of E. coli were determined by micro-broth dilution method. The fractional inhibitory concentration( FIC) of ARS combining with 7 antibiotics against resistant E.coli was determined by checkerboard dilution method. The plate perforation method was used to study the effect of ARS pretreatment on the diameter of antibiotic inhibition zone. ARS had no obvious antibacterial effect when used alone. However,when combined with antibiotics,ARS( ≥512 μg/m L) could significantly reduce the MIC value of antibiotics. The FIC indexes of6 antibiotics were all less than 0.5 except for streptomycin,showing synergistic effects. In addition,pretreatment by 512 μg/m L and 1024 μg/m L ARS increased the inhibition zone diameter of both streptomycin and colistin against resistant E. coli, with no effects to other tested antibiotics. Taken together, ARS has no inherent antibacterial activity,but it can significantly enhance the antibacterial effect of antibiotics against drug-resistant E.coli.
引文
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[4] Wang J,Zhou H,Zheng J,et al. The antimalarial artemisinin synergizes with antibiotics to protect against lethal live Escherichia coli challenge by decreasing proinflammatory cytokine release[J].Antimicrobial Agents and Chemotherapy,2006,50(7):2420-2427.
[5]金慧玲,张汝芝,高玉祥,等.青蒿琥酯抗真菌、抗细菌的实验研究[J].中国微生态学杂志,2003,15(1):26-28.Jin H L,Zhang Y Z,Gao Y X,et al. Experimental Study on Antifungal and Antibacterial Activity of Artesunate[J]. Journal of Chinese Microecology,2003,15(1):26-28.
[6] Li B,Yao Q,Pan XC,Wang N,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-Tol C[J].Journal of Antimicrobial Chemotherapy,2011; 66(4):769-77.
[7]李琛琛,尹昆,李敏,等.青蒿琥酯联合头孢曲松钠对铜绿假单胞菌抗菌作用研究[J].中国病原生物学杂志,2015,10(10):883-887.Li C C,Yin K,Li M,et al. Study on the antibacterial activity of artesunate in combination with ceftriaxone sodium[J]. Journal of Pathogen Biology,2015,10(10):883-887.
[8] Brinda C,Sundar P,Kaliappan I,et al. Inhibition of New Delhi Metallo-β-Lactamase 1(NDM-1)Producing Escherichia coli IR-6 by Selected Plant Extracts and Their Synergistic Actions with Antibiotics[J]. Frontiers in Microbiology,2017,8:1580.
[9] Antonio C,Silvia P,Colavecchio O L,et al. Synergistic Activity of Colistin in Combination With Resveratrol Against ColistinResistant Gram-Negative Pathogens[J]. Frontiers in Microbiology,2018,9:1808.
[10] Salem A Z M,Khusro A,Elghandour M M Y,et al. Susceptibility of ruminal bacteria isolated from large and small ruminant to multiple conventional antibiotics[J]. Microbial Pathogenesis,2018,121:93-99.
[11] Ling L L,Schneider T,Peoples A J,et al. Erratum:A new antibiotic kills pathogens without detectable resistance[J]. Nature,2015,520(7547):388-388.
[12] G20 Leaders'Communique Hangzhou Summit[J]. China Economic Weekly,2016,(36):98-105.
[13] Brown,David. Antibiotic resistance breakers:can repurposed drugs fill the antibiotic discovery void?[J]. Nature Reviews Drug Discovery,2015,14(12):821–832.
[14] Lin A X,Chan G,Hu Y,et al. Internationalization of traditional Chinese medicine:Current international market,internationalization challenges and prospective suggestions[J]. Chinese Medicine,2018,13(1):9.
[15] Ho W E,Peh H Y,Chan T K,et al. Artemisinins:Pharmacological actions beyond anti-malarial[J]. Pharmacology&Therapeutics,2014,142(1):126-139.
[16] Feng F B,Qiu H Y. Effects of Artesunate on chondrocyte proliferation,apoptosis and autophagy through the PI3K/AKT/m TOR signaling pathway in rat models with rheumatoid arthritis[J].Biomedicine&Pharmacotherapy,2018,102:1209-1220.