萘酚喹与阿奇霉素伍用延缓恶性疟原虫对萘酚喹的抗性
|
摘要
目的观察实验组恶性疟原虫经萘酚喹与阿奇霉素伍用与对照组恶性疟原虫经萘酚喹单独用药在药物作用下体外敏感性的变化,评价萘酚喹与阿奇霉素伍用对延缓恶性疟原虫抗性的作用。方法通过体外药物剂量递增间隔作用法对实验室驯化的恶性疟原虫FccSM/YN株进行长期抗性培育,采用Rieckmann微量测定法,在不同时段(d)测定疟原虫的体外敏感性,用ICEstimator软件计算半数抑制浓度(IC_(50)),药物作用后实验组、对照组的IC_(50)与亲代相比,计算抗性指数,抗性指数<2,判定为敏感,抗性指数≥2为抗性。结果药物作用前,萘酚喹对恶性疟原虫IC_(50)值为2.11 nmol/L,阿奇霉素为3.21 nmol/L。对照组经萘酚喹作用120 d后,萘酚喹IC_(50)值升至65.47 nmol/L,阿奇霉素上升至68.58 nmol/L,萘酚喹/阿奇霉素分别是10.34 nmol/L,抗性指数均>2。实验组经萘酚喹/阿奇霉素作用120 d后,萘酚喹对恶性疟原虫的IC_(50)值升至35.47 nmol/L,阿奇霉素上升至50.04 nmol/L,抗性指数均>2。萘酚喹/阿奇霉素IC_(50)值分别为2.89 nmol/L,抗性指数<2。实验组与对照组相比,实验组在同一药物浓度的疟原虫裂殖体抑制率明显高于对照组(P<0.01);实验组IC_(50)值明显低于对照组;萘酚喹/阿奇霉素的抗性指数<2。结论本研究方法可以培育出恶性疟原虫萘酚喹抗性株;萘酚喹与阿奇霉素伍用可延缓恶性疟原虫对萘酚喹和萘酚喹/阿奇霉素抗性的产生与发展。
Objective To evaluate the effect of combination of naphthoquine and azithromycin in slowing down naphthoquine resistance by in vitro testing naphthoquine susceptibility of Plasmodium falciparum in the experimental group(combination of naphthoquine and azithromycin) and control group(naphthoquine alone). Methods The domesticated P.falciparum FccSM/YN strain was cultured to develop resistance by using in vitro drug dose-increasing interval method for a long-term. The in vitro sensitivity of Plasmodium was determined by Rieckmann microassay at different time points(d).ICEstimator software was used to analyse the results. The half-inhibitory concentration of naphthalene against Plasmodium falciparum(IC_(50)) was calculated by software. After the drug was added, the IC_(50) of the experimental group and the control group was compared with the parental value, and the resistance index was calculated. The resistance index< 2 was judged as sensitive, and vice versa. Results The IC_(50) was 2.11 nmol/L for naphthalene and 3.21 nmol/L for azithromycin before drug treatment, respectively. In control group, after 120 days of naphthalene treatment, the IC_(50) value of naphthalene increased to65.47 nmol/L, that of azithromycin increased to 68.58 nmol/L, that of naphthalene/azithromycin was 10.34 nmol/L, so all the resistance indexes were >2. In the experimental group, the IC_(50) value of naphthalene increased to 35.47 nmol/L, and that of azithromycin increased to 50.04 nmol/L, and all the two resistance indexes were >2, however, the IC_(50) value of naphthalene/azithromycin was 2.89 nmol/L and the resistance index was <2. Compared with the control group, the inhibition rate of Plasmodium schizont in the experimental group was significantly higher than that in the control group(P<0.01). The IC_(50) value of the experimental group was significantly lower than that of the control group; the resistance index of naphthalene/azithromycin was <2. Conclusion This study in vitro can culture a naphtholquine-resistant strain of Plasmodium falciparum.The combination of naphthoquine and azithromycin can slow down the development of naphthoquine and naphthoquine/azimycin resistance in Plasmodium falciparum.
Objective To evaluate the effect of combination of naphthoquine and azithromycin in slowing down naphthoquine resistance by in vitro testing naphthoquine susceptibility of Plasmodium falciparum in the experimental group(combination of naphthoquine and azithromycin) and control group(naphthoquine alone). Methods The domesticated P.falciparum FccSM/YN strain was cultured to develop resistance by using in vitro drug dose-increasing interval method for a long-term. The in vitro sensitivity of Plasmodium was determined by Rieckmann microassay at different time points(d).ICEstimator software was used to analyse the results. The half-inhibitory concentration of naphthalene against Plasmodium falciparum(IC_(50)) was calculated by software. After the drug was added, the IC_(50) of the experimental group and the control group was compared with the parental value, and the resistance index was calculated. The resistance index< 2 was judged as sensitive, and vice versa. Results The IC_(50) was 2.11 nmol/L for naphthalene and 3.21 nmol/L for azithromycin before drug treatment, respectively. In control group, after 120 days of naphthalene treatment, the IC_(50) value of naphthalene increased to65.47 nmol/L, that of azithromycin increased to 68.58 nmol/L, that of naphthalene/azithromycin was 10.34 nmol/L, so all the resistance indexes were >2. In the experimental group, the IC_(50) value of naphthalene increased to 35.47 nmol/L, and that of azithromycin increased to 50.04 nmol/L, and all the two resistance indexes were >2, however, the IC_(50) value of naphthalene/azithromycin was 2.89 nmol/L and the resistance index was <2. Compared with the control group, the inhibition rate of Plasmodium schizont in the experimental group was significantly higher than that in the control group(P<0.01). The IC_(50) value of the experimental group was significantly lower than that of the control group; the resistance index of naphthalene/azithromycin was <2. Conclusion This study in vitro can culture a naphtholquine-resistant strain of Plasmodium falciparum.The combination of naphthoquine and azithromycin can slow down the development of naphthoquine and naphthoquine/azimycin resistance in Plasmodium falciparum.
引文
[1]WHO.The WHO Global Malaria Programme.Word malaria report2017[R].Switzerland:World Health Organization,2018.
[2]丰俊,张丽,张少森,等.全国2005-2015年疟疾疫情分析[J].中国热带医学,2017,17(4):325-335.
[3]杨恒林,肖宁,杨亚明,等.中缅和中老边境地区消除疟疾的挑战、机遇与对策[J].中国热带医学,2017,17(4):321-324,335.
[4]高琪.我国消除疟疾面临的机遇与挑战[J].中国血吸虫病防治杂志,2011,23(4):347-349.
[5]汤林华,TANG Lin-hua.中国消除疟疾的目标、策略与路径[J].中国热带医学,2016,16(4):301-304.
[6]YANG H L,YANG Y M,YANG P F,et al.Monitoring Plasmodium falciparum chloroquine resistance in Yunnan Province,China,1981-2006[J].Acta Trop,2008,108(1):44-49.
[7]SáJ M,CHONGJ L,WELLEMST E.Malaria drug resistance:New observations and developments[J].Essays Biochem,2011,51:137-160.
[8]燕贺,丰俊,尹建海,等.不同地理来源恶性疟原虫微卫星标记位点多态性研究[J].中国血吸虫病防治杂志,2018,30(5):504-507.
[9]陈静,张梅花,唐建霞,等.非恶性疟人体疟原虫药物抗性分子标记研究进展[J].中国血吸虫病防治杂志,2018,30(4):465-471.
[10]WONGSRICHANALAI C.Artemisinin resistance or artemisininbased combination therapy resistance?[J].Lancet Infect Dis,2013,13(2):114-115.
[11]赵绍敏,王满元.恶性疟原虫对青蒿素类药物产生耐药性的全球现状和基础研究[J].中国寄生虫学与寄生虫病杂志,2014,32(5):380-384.
[12]孙艾明,董莹,陈梦妮,等.云南省恶性疟原虫青蒿素耐药性相关基因K13kelch结构域序列多态性的分析[J].中国寄生虫学与寄生虫病杂志,2016,34(4):339-345.
[13]刘克辛.药理学[M].北京:清华大学出版社,2012.
[14]KUSCHNERR A,HEPPNERD G,ANDERSENS L,et al.Azithromycin prophylaxis against a chloroquine-resistant strain of Plasmodium falciparum[J].Lancet,1994,343(8910):1396-1397.
[15]王家文,韦明标,廖豪峰,等.阿奇霉素治疗或预防疟疾的研究进展[J].热带医学杂志,2017,17(1):129-132,137.
[16]TAYLORW R,RICHIET L,FRYAUFFD J,et al.Tolerability of azithromycin as malaria prophylaxis in adults in northeast papua,indonesia[J].Antimicrob Agents Chemother,2003,47(7):2199-2203.
[17]李豫,陈华,杨恒林.萘酚喹单用及其与阿奇霉素伍用体外抑制恶性疟原虫作用研究[J].中国病原生物学杂志,2011,6(7):505-506,520.
[18]王京燕,李国福,赵京花,等.磷酸萘酚喹与青蒿素伍用增效和延缓疟原虫抗药性的研究[J].寄生虫与医学昆虫学报,2008,15(3):133-136.
[19]YANG H L,WANGJ Y,LIU H,et al.Malaria prophylaxis using naphthoquine-azithromycin combination:randomized,doubleblind,placebo-controlled studies to assess safety and protective efficacy in Southeast Asia[J].Antimicrob Agents Chemother,2018,62(9):e00793-18.
[20]孙院红,周家莲,刘慧,等.云南省恶性疟原虫抗萘酚喹株的体外培育[J].中国病原生物学杂志,2010,5(2):124-126,133.
[21]高白荷,杨恒林,黄开国,等.云南省恶性疟原虫抗青蒿琥酯株的体外培育[J].中国寄生虫病防治杂志,2000,13(3):215-216.
[22]RIECKMANNK H,CAMPBELLG H,SAXL J,et al.Drug sensitivity of Plasmodium falciparum.An in-vitro microtechnique[J].Lancet,1978,1(8054):22-23.
[23]KADDOURI H,NAKACHE S,HOUZéS,et al.Assessment of the drug susceptibility of Plasmodium falciparum clinical isolates from africa by using a Plasmodium lactate dehydrogenase immunodetection assay and an inhibitory maximum effect model for precise measurement of the 50-percent inhibitory concentration[J].Antimicrob Agents Chemother,2006,50(10):3343-3349.2019-03-28
[2]丰俊,张丽,张少森,等.全国2005-2015年疟疾疫情分析[J].中国热带医学,2017,17(4):325-335.
[3]杨恒林,肖宁,杨亚明,等.中缅和中老边境地区消除疟疾的挑战、机遇与对策[J].中国热带医学,2017,17(4):321-324,335.
[4]高琪.我国消除疟疾面临的机遇与挑战[J].中国血吸虫病防治杂志,2011,23(4):347-349.
[5]汤林华,TANG Lin-hua.中国消除疟疾的目标、策略与路径[J].中国热带医学,2016,16(4):301-304.
[6]YANG H L,YANG Y M,YANG P F,et al.Monitoring Plasmodium falciparum chloroquine resistance in Yunnan Province,China,1981-2006[J].Acta Trop,2008,108(1):44-49.
[7]SáJ M,CHONGJ L,WELLEMST E.Malaria drug resistance:New observations and developments[J].Essays Biochem,2011,51:137-160.
[8]燕贺,丰俊,尹建海,等.不同地理来源恶性疟原虫微卫星标记位点多态性研究[J].中国血吸虫病防治杂志,2018,30(5):504-507.
[9]陈静,张梅花,唐建霞,等.非恶性疟人体疟原虫药物抗性分子标记研究进展[J].中国血吸虫病防治杂志,2018,30(4):465-471.
[10]WONGSRICHANALAI C.Artemisinin resistance or artemisininbased combination therapy resistance?[J].Lancet Infect Dis,2013,13(2):114-115.
[11]赵绍敏,王满元.恶性疟原虫对青蒿素类药物产生耐药性的全球现状和基础研究[J].中国寄生虫学与寄生虫病杂志,2014,32(5):380-384.
[12]孙艾明,董莹,陈梦妮,等.云南省恶性疟原虫青蒿素耐药性相关基因K13kelch结构域序列多态性的分析[J].中国寄生虫学与寄生虫病杂志,2016,34(4):339-345.
[13]刘克辛.药理学[M].北京:清华大学出版社,2012.
[14]KUSCHNERR A,HEPPNERD G,ANDERSENS L,et al.Azithromycin prophylaxis against a chloroquine-resistant strain of Plasmodium falciparum[J].Lancet,1994,343(8910):1396-1397.
[15]王家文,韦明标,廖豪峰,等.阿奇霉素治疗或预防疟疾的研究进展[J].热带医学杂志,2017,17(1):129-132,137.
[16]TAYLORW R,RICHIET L,FRYAUFFD J,et al.Tolerability of azithromycin as malaria prophylaxis in adults in northeast papua,indonesia[J].Antimicrob Agents Chemother,2003,47(7):2199-2203.
[17]李豫,陈华,杨恒林.萘酚喹单用及其与阿奇霉素伍用体外抑制恶性疟原虫作用研究[J].中国病原生物学杂志,2011,6(7):505-506,520.
[18]王京燕,李国福,赵京花,等.磷酸萘酚喹与青蒿素伍用增效和延缓疟原虫抗药性的研究[J].寄生虫与医学昆虫学报,2008,15(3):133-136.
[19]YANG H L,WANGJ Y,LIU H,et al.Malaria prophylaxis using naphthoquine-azithromycin combination:randomized,doubleblind,placebo-controlled studies to assess safety and protective efficacy in Southeast Asia[J].Antimicrob Agents Chemother,2018,62(9):e00793-18.
[20]孙院红,周家莲,刘慧,等.云南省恶性疟原虫抗萘酚喹株的体外培育[J].中国病原生物学杂志,2010,5(2):124-126,133.
[21]高白荷,杨恒林,黄开国,等.云南省恶性疟原虫抗青蒿琥酯株的体外培育[J].中国寄生虫病防治杂志,2000,13(3):215-216.
[22]RIECKMANNK H,CAMPBELLG H,SAXL J,et al.Drug sensitivity of Plasmodium falciparum.An in-vitro microtechnique[J].Lancet,1978,1(8054):22-23.
[23]KADDOURI H,NAKACHE S,HOUZéS,et al.Assessment of the drug susceptibility of Plasmodium falciparum clinical isolates from africa by using a Plasmodium lactate dehydrogenase immunodetection assay and an inhibitory maximum effect model for precise measurement of the 50-percent inhibitory concentration[J].Antimicrob Agents Chemother,2006,50(10):3343-3349.2019-03-28