目的 考察热毒宁注射液对EV71病毒感染Vero细胞及乳鼠致手足口病模型的治疗作用及其机制。方法 体外模型采用33 TCID₅₀和25 TCID₅₀ EV71感染Vero细胞，建立细胞感染病毒模型；采用乳鼠ip EV71病毒100 μL/只的方法制备体内模型，攻毒2 h后各组分别ip给予相应药物，观察热毒宁注射液（2.60、1.30、0.65 g/kg）和阳性对照药利巴韦林（10 mg/kg）对乳鼠体质量、临床评分和肌肉组织中白细胞介素-1β（IL-1β）、IL-6、肿瘤坏死因子-α（TNF-α）和单核细胞趋化蛋白-1（MCP-1）水平及腿部肌肉病理改变的影响。结果 体外结果显示，在Vero细胞模型上，热毒宁注射液（生药5.0 mg/mL）显著抑制33 TCID₅₀和25 TCID₅₀ EV71引起的细胞病变；体内实验结果显示，与模型组比较，高、中剂量热毒宁注射液可降低病毒感染乳鼠的临床评分，提高乳鼠体质量，改善乳鼠生存状态，降低乳鼠肌肉组织中细胞因子IL-1β、IL-6、TNF-α、MCP-1水平（P<0.05、0.01）；与模型组比较，热毒宁注射液可显著改善肌肉组织病理学病变。结论 热毒宁注射液能够抑制EV71病毒导致的乳鼠死亡，明显延长乳鼠的存活时间，能够减轻病毒引起的临床症状，缓解病毒引发的乳鼠生长抑制，抑制IL-1β、IL-6、TNF-α、MCP-1水平可能是其治疗手足口病的重要机制。

Objective To investigate the effect of Reduning Injection on Vero cells and suckling mice infected by EV71 virus and its mechanism in vitro and in vivo.Methods The anti-virus ability was exhibited in Vero cells which were infected by 33 TCID50 and 25 TCID50 titers of EV71 in vitro.Meanwhile, the antiviral activity in vivo was evaluated on suckling mice model, in which suckling mice were intraperitoneal injected with EV71 at 100 μL.After 2 h infection, Reduning Injection 2.60, 1.30, and 0.65 g/kg and ribavirin 10 mg/kg were followed for treatment.To further understand the antiviral effects of Reduning Injection, weigh of suckling mice, clinical symptoms, expression of IL-1β, IL-6, MCP-1, and TNF-α in skeletal muscle were recorded.Moreover, histological examination of skeletal muscle was examined.Results The results demonstrated that Reduning Injection 5.0 mg/mL could obviously inhibit Vero cells lesion induced by EV71 in vitro.Compared with the model group, suckling mouse model results showed the high and middle dose of Reduning Injection could reduce clinical manifestations scores, increase the weight and improve living conditions of model mice, decrease the expression of IL-1β, IL-6, TNF-α, and MCP-1 in skeletal muscle of suckling mice (P<0.05, 0.01).Histological damages in skeletal muscle were also alleviated with Reduning Injection treatment.Conclusion In a suckling mouse model, Reduning Injection can reduce mortality and obviously prolong the survival time of suckling mice caused by EV71 infection.It can alleviate clinical symptoms and growth inhibition caused by the EV71 virus.Inhibition of IL-1β, IL-6, TNF-α, and MCP-1 levels may be the important mechanism of Reduning Injection in the treatment of hand-foot-and-mouth disease.

科技部重大新药创制：现代中药创新集群与数字制药技术平台（2013ZX09402203）