手足清栓通过抗病毒降低71型肠道病毒(EV71)感染小鼠死亡率

Shouzuqing suppository can reduce the mortality of EV71 infected suckling mice by antiviral effect

71型肠道病毒(enterovirus 71,EV71)是引起手足口病(hand,foot and mouth disease,HFMD)的主要病原体,在亚太地区流行,至今尚无特效药物。手足清栓作为治疗手足口病的中药方剂因可接受性强、无可见副作用而受到青睐,但治疗效果和作用机理至今尚未见有报道。为阐明手足清栓对手足口病模型的治疗效果和作用机理,本研究测定手足清栓对健康Vero细胞的毒性和对EV71-BJ株感染Vero细胞模型的治疗效果,测定药物作用与否的Vero细胞感染模型的病毒载量;测定手足清栓经灌肠途径对健康小鼠的毒性和对EV71-BJ株感染小鼠模型的治疗效果,重点观察手足清栓药物对EV71-BJ株重症感染小鼠模型死亡率、体质量、临床症状的改善情况,测定手足清栓经灌肠途径给药对EV71-BJ株重症感染小鼠模型靶组织产生的病变程度、病毒载量、炎性分子的变化特征,从而探索手足清栓治疗EV71感染的机制。体外结果显示,在EV71-BJ株感染Vero细胞模型上,于对细胞无肉眼可见毒性质量浓度下(生药60 mg/L),手足清栓通过降低感染细胞的病毒载量显著抑制EV71感染引起的Vero细胞病变;体内结果显示,在无毒质量浓度条件下(生药11.05 g/kg),手足清栓通过降低病毒载量、改善组织病理变化、降低炎性分子浓度机制降低EV71感染小鼠模型的死亡率,改善发病小鼠的临床症状。结果表明,手足清栓可通过抗病毒明显提高小鼠的存活率,能够减轻EV71感染引起的HFMD的临床症状。

Enterovirus 71(EV71)is the main pathogen that causes hand,foot and mouth disease(HFMD).As a first-line traditional Chinese medicine prescription for the treatment of HFMD,Shouzuqing suppository is favored by infants in Beijing due to its strong acceptability and no visible side effects,but the therapeutic effect and mechanism has not been clarified so far.To clarify the therapeutic effect and mechanism of Shouzuqing suppository on HFMD model.The toxicity of Shouzuqing suppository on Vero cells and the therapeutic effect on the Vero cell model infected by EV71-BJ strain were measured.Determination of the toxicity to healthy mice and the treatment effect on the EV71-BJ strains infected model were explored.Survival rates,body weight,the improvement of clinical symptoms were determined.The target tissues lesion severity,viral loads and the concentrations of inflammatory molecules were explored to clarify the treatment mechanism.In vitro results showed that on the Vero cell model infected by EV71-BJ strain,Shouzuqing suppository significantly reduced Vero cell cytopathic effect caused by EV71 infection by inhibiting the viral multiplication at the concentration of 60 mg/L.In vivo results showed that under the condition of non-toxic concentration(raw drug 11.05 g/kg),Shouzuqing suppository reduced the mortality rate of EV71 infected suckling mouse model and improved the clinical symptoms by inhibiting the viral load,improving the tissues pathology changes and reducing the inflammatory molecular concentration.Shouzuqing suppository can significantly improve the survival rate of suckling mice through anti-viral effect.