小儿热速清糖浆防治手足口病的体内外药效学研究
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摘要
手足口病(Hand,foot and mouth disease,HFMD)是由肠道病毒感染引起的一种儿童常见传染病,全球普遍高发,严重威胁患儿健康。目前缺乏有效治疗药物,主要进行对症治疗,因此,研发高效的抗HFMD相关肠道病毒的药物和治疗方法十分重要。本研究应用细胞和小动物感染模型评价了小儿热速清糖浆(RSQ)对肠道病毒A组71型(Enterovirus A71,EV-A71)致病能力的抑制作用。体内实验中,小儿热速清糖浆设计66ml/(kg·d~(-1))、33ml/(kg·d~(-1))、16.5ml/(kg·d~(-1))三个剂量组,分别对EV-A71 H株感染的BALB/c乳鼠进行预防性和治疗性给药,观察乳鼠感染的严重程度及死亡数、动物死亡率、死亡保护率以及生命延长率。体外实验中,采用CPE法观察小儿热速清糖浆对EV-A71 H株、EV-A71 BrCr株、柯萨奇病毒(Coxsackievirus,Cox)B_3株、Cox B_4株、Cox B_5株所致细胞病变的抑制作用。体内试验结果显示,预防性和治疗性给予小儿热速清糖浆后,中、小剂量组乳鼠的感染程度积分及死亡率均明显降低,并可以延长动物的存活时间,大剂量组未表现出明显的药效作用。体外实验结果显示,小儿热速清糖浆对EV-A71 H株、Cox B_5株、EV-A71 BrCr株及Cox B_3株所致RD或Hep-2细胞病变具有一定的抑制作用,治疗指数分别为5.77、5.01、3.98、2.30。以上结果表明,小儿热速清糖浆对EV-A71 H株感染的手足口病乳鼠模型具有显著的预防和治疗作用,在体外对手足口病致病病毒EV-A71 H株、EV-A71 BrCr株、Cox B_3及Cox B_5株均有明显的抑制作用。
Hand,foot and mouth disease(HFMD) was a common infectious disease caused by enterovirus infection. It was widespread in the world and threatened the health of children. At present,there was no effective treatments,so it was important to develop high-efficiency drugs and treatments related to HFMD. In this study,the cell and small animal infection models were used to evaluate the inhibitory effect of Xiaoer Resuqing syrup(RSQ)on the pathogenicity of Enterovirus A71(EV-A71). In the experiment in vivo,three dose groups(66,33,16.5 ml/(kg·d~(-1)))were designed. Prophylactic and therapeutic administration of RSQ was applied to mice infected with EV-A71 H strain. We observed the degree of infection,mortality and survival.In the experiment in vitro,inhibition by RSQ at five non-toxic doses of EV-A71 H as well as EV-A71 Brcr, Coxsackievirus(Cox) B3,Cox B_4 and Cox B_5 strains was determined by the cytopathogenic-effect method on Hep-2 or RD cells. The results showed that medium and low-dose groups of RSQ could reduce the degree of infection and mortality,and simultaneously increase survival significantly. RSQ showed significant preventive and therapeutic effects on suckling BALB/c mice infected with EV-A71 H strain. At no-toxic doses,RSQ inhibited infection with EV-A71 H as well as that by EV-A71 Brcr,Cox B_3 and Cox B_5 strains visibly,and the therapeutic index was 5.77,5.01,3.98 and 2.30,respectively. The results indicated that RSQ has a good pharmacologic effect against HFMD due to EV-A71 H strain infection in vivo,and has strong effects against HFMD due to infection with EV-A71 H,EV-A71 BrCr,Cox B_5 and Cox B_3 strains in vitro.
Hand,foot and mouth disease(HFMD) was a common infectious disease caused by enterovirus infection. It was widespread in the world and threatened the health of children. At present,there was no effective treatments,so it was important to develop high-efficiency drugs and treatments related to HFMD. In this study,the cell and small animal infection models were used to evaluate the inhibitory effect of Xiaoer Resuqing syrup(RSQ)on the pathogenicity of Enterovirus A71(EV-A71). In the experiment in vivo,three dose groups(66,33,16.5 ml/(kg·d~(-1)))were designed. Prophylactic and therapeutic administration of RSQ was applied to mice infected with EV-A71 H strain. We observed the degree of infection,mortality and survival.In the experiment in vitro,inhibition by RSQ at five non-toxic doses of EV-A71 H as well as EV-A71 Brcr, Coxsackievirus(Cox) B3,Cox B_4 and Cox B_5 strains was determined by the cytopathogenic-effect method on Hep-2 or RD cells. The results showed that medium and low-dose groups of RSQ could reduce the degree of infection and mortality,and simultaneously increase survival significantly. RSQ showed significant preventive and therapeutic effects on suckling BALB/c mice infected with EV-A71 H strain. At no-toxic doses,RSQ inhibited infection with EV-A71 H as well as that by EV-A71 Brcr,Cox B_3 and Cox B_5 strains visibly,and the therapeutic index was 5.77,5.01,3.98 and 2.30,respectively. The results indicated that RSQ has a good pharmacologic effect against HFMD due to EV-A71 H strain infection in vivo,and has strong effects against HFMD due to infection with EV-A71 H,EV-A71 BrCr,Cox B_5 and Cox B_3 strains in vitro.
引文
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[5]王敏,陈荣昌,季宇彬,韩冰,葛一蒙,王姗,孙桂波,孙晓波.小儿热速清糖浆抗流感病毒和抗菌作用研究[J].中国比较医学杂志,2018,28(8):83-89.
[6]郭姗姗,金亚宏,王意忠,高英杰,时宇静,马荣,刘方舟,钟菊迎,田雪川,崔晓兰.疏风解毒颗粒防治手足口病研究的体内外药效学研究[J].中国实验方剂学杂志,2012,18(2):206-209.
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[8]韩淑娴,陈影,张倩,韩冰,葛一蒙,项彦华,廖福龙,游云.流动条件下血栓通胶囊抗血小板粘附的分子药理学机制研究[J].中国中药杂志,2017,42(2):341-346.
[9]于长水.天津市由CoxA16肠道病毒引起的手足口病流行[J].中华流行病学杂志,1985,6(2):66-69.
[10]Li W,Yi L,Su J,Lu J,Ke CW,Zeng H R,Guan DW,Ma C,Zhang W L,Xiao H,Li H,Lin J Y,Zhang Y H.Seroprevalence of human enterovirus 71 and coxsackievirus A16 in Guangdong,China,in pre-and post-2010 HFMD epidemic period[J/OL].PLoS One,2013,8(12):e80515.
[11]Wang Y R,Sun L L,Xiao W L,Chen L Y,Wang XF,Pan D M.Epidemiology and clinical characteristics of hand,foot,and mouth disease in a Shenzhen sentinel hospital from 2009 to 2011[J].BMC Infect Dis,2013,13:539.
[12]Xu M,Su L,Cao L,Zhong H,Dong N,Xu J.Enterovirus genotypes causing hand,foot,and mouth disease in Shanghai,China:a molecular epidemiological analysis[J].BMC Infect Dis,2013,13:489.
[13]Ni H X,Yi B,Yin J H,Fang T,He T F,Du Y,Wang J,Zhang H W,Xie L,Ding Y B,Gu W Z,Zhang S,Han Y F,Dong H J,Su T,Xu G Z,Cao GW.Epidemiological and etiological characteristics of hand,foot,and mouth disease in Ningbo,China,2008-2011[J].J Clin Virol,2012,54(4):342-348.
[14]Du J,Wang X,Hu Y,Li Z,Li Y,Sun S,Yang F,Jin Q.Changing aetiology of hand,foot,and mouth disease in Linyi,China,2009-2011[J].Clin Microbiol Infect,2014,20(1):O47-O49.
[15]Zhang Y,Tan X,Cui A,Mao N,Xu S,Zhu S,Zhou J,Shi J,Zhao Y,Wang X,Huang X,Zhu S,Zhang Y,Tang W,Ling H,Xu W.Complete genome analysis of the C4 subgenotype strains of enterovirus 71:predominant recombination C4 viruses presistently circulating in China for 14 years[J/OL].PLoS One,2013,8(2):e56341.
[16]张斐,杜武勋,朱明丹,许艳伶,冯利民.中药复方量效关系研究现状[J].中医杂志,2013,54(1):74-77.
[17]傅延齡,蔡坤坐,宋佳.方药量效关系文献与理论研究思考[J].北京中医药大学学报,2010,33(9):601-609.
[18]郭姗姗,马雪萍,潘昕,高英杰,崔晓兰.复方一枝蒿颗粒对防治手足口病的体内外药效学研究[J].中国药物警戒,2014,11(7):385-389.
[2]Chang G H,Luo Y J,Wu X Y,Si B Y,Lin L,Zhu QY.Monoclonal antibody induced with inactived EV71-Hn2 virus protects mice against lethal EV71-Hn2 virus infection[J].Virol J,2010,7:106.
[3]Lin Y W,Chang K C,Kao C M,Chang S P,Tung YY,Chen S H.Lymphocyte and antibody responses reduce enterovirus 71 lethality in mice by decreasing tissue viral loads[J].J Virol,2009,83(13):6477-6483.
[4]潘红春,王海英,彭苍娇,管敏唱.小儿热速清糖浆治疗婴幼儿上呼吸道感染疗效观察[J].海峡药学,2013,25(8):126-127.
[5]王敏,陈荣昌,季宇彬,韩冰,葛一蒙,王姗,孙桂波,孙晓波.小儿热速清糖浆抗流感病毒和抗菌作用研究[J].中国比较医学杂志,2018,28(8):83-89.
[6]郭姗姗,金亚宏,王意忠,高英杰,时宇静,马荣,刘方舟,钟菊迎,田雪川,崔晓兰.疏风解毒颗粒防治手足口病研究的体内外药效学研究[J].中国实验方剂学杂志,2012,18(2):206-209.
[7]王意忠,崔晓兰,高英杰,郭姗姗,王秀坤,黄洋,赵晔,巩文峰.栀子提取物抗病毒试验研究[J].中国中药杂志,2006,31(14):1176-1178.
[8]韩淑娴,陈影,张倩,韩冰,葛一蒙,项彦华,廖福龙,游云.流动条件下血栓通胶囊抗血小板粘附的分子药理学机制研究[J].中国中药杂志,2017,42(2):341-346.
[9]于长水.天津市由CoxA16肠道病毒引起的手足口病流行[J].中华流行病学杂志,1985,6(2):66-69.
[10]Li W,Yi L,Su J,Lu J,Ke CW,Zeng H R,Guan DW,Ma C,Zhang W L,Xiao H,Li H,Lin J Y,Zhang Y H.Seroprevalence of human enterovirus 71 and coxsackievirus A16 in Guangdong,China,in pre-and post-2010 HFMD epidemic period[J/OL].PLoS One,2013,8(12):e80515.
[11]Wang Y R,Sun L L,Xiao W L,Chen L Y,Wang XF,Pan D M.Epidemiology and clinical characteristics of hand,foot,and mouth disease in a Shenzhen sentinel hospital from 2009 to 2011[J].BMC Infect Dis,2013,13:539.
[12]Xu M,Su L,Cao L,Zhong H,Dong N,Xu J.Enterovirus genotypes causing hand,foot,and mouth disease in Shanghai,China:a molecular epidemiological analysis[J].BMC Infect Dis,2013,13:489.
[13]Ni H X,Yi B,Yin J H,Fang T,He T F,Du Y,Wang J,Zhang H W,Xie L,Ding Y B,Gu W Z,Zhang S,Han Y F,Dong H J,Su T,Xu G Z,Cao GW.Epidemiological and etiological characteristics of hand,foot,and mouth disease in Ningbo,China,2008-2011[J].J Clin Virol,2012,54(4):342-348.
[14]Du J,Wang X,Hu Y,Li Z,Li Y,Sun S,Yang F,Jin Q.Changing aetiology of hand,foot,and mouth disease in Linyi,China,2009-2011[J].Clin Microbiol Infect,2014,20(1):O47-O49.
[15]Zhang Y,Tan X,Cui A,Mao N,Xu S,Zhu S,Zhou J,Shi J,Zhao Y,Wang X,Huang X,Zhu S,Zhang Y,Tang W,Ling H,Xu W.Complete genome analysis of the C4 subgenotype strains of enterovirus 71:predominant recombination C4 viruses presistently circulating in China for 14 years[J/OL].PLoS One,2013,8(2):e56341.
[16]张斐,杜武勋,朱明丹,许艳伶,冯利民.中药复方量效关系研究现状[J].中医杂志,2013,54(1):74-77.
[17]傅延齡,蔡坤坐,宋佳.方药量效关系文献与理论研究思考[J].北京中医药大学学报,2010,33(9):601-609.
[18]郭姗姗,马雪萍,潘昕,高英杰,崔晓兰.复方一枝蒿颗粒对防治手足口病的体内外药效学研究[J].中国药物警戒,2014,11(7):385-389.