香菊感冒颗粒由香薷、藿香和野菊花等多味中药组成，对风寒、风热及寒包火三类证型的感冒均有较好的退热效果。在体外用CCK8试剂盒检测香菊感冒颗粒的细胞毒性，及其对甲型流感病毒(H1N1、H3N2、H1N1奥司他韦耐药株)和其他呼吸道病毒(RSV、仙台病毒和腺病毒)感染细胞的保护作用，且观察病毒感染后药物治疗对细胞病变效应(Cytopathogenic effect，CPE)的影响。在体内建立H1N1病毒感染小鼠模型，评价药物治疗后小鼠生存指标，通过HE染色、ELISA、Westernblot和免疫组化分析药物治疗对小鼠炎症因子释放及炎症信号通路活化的影响。结果显示，香菊感冒颗粒的半数细胞毒浓度(Concentration of cytotoxicity 50％，CC50)高于其对受试病毒株的有效抑制浓度，提示其具有良好的安全性。针对甲型流感病毒及仙台病毒引起的MDCK细胞病变显示出良好的抑制效果，尤其对H1N1奥司他韦耐药株也显示出了一定的抑制活性。香菊感冒颗粒在体内能够明显降低H1N1病毒感染小鼠的死亡率，延长其存活时间，并能降低肺指数，缓解肺部炎性细胞浸润。同时小鼠肺部炎症因子释放、NLRP3-caspase1-IL-1β蛋白的活化也明显下降。研究通过体外建立6种病毒感染的细胞模型，体内建立H1N1病毒感染的小鼠模型，证明了香菊感冒颗粒具有良好的体内外抗病毒活性，并可以通过抑制炎症信号NLRP3-caspase1-IL-1β通路抑制由H1N1病毒感染引起的肺炎。
Xiangju cold granules is composed of Chinese herbs such as Chinese Mosla,patchouli and wild chrysanthemum.It has good antipyretic effect on cold,wind heat and cold wrapped fire.The cytotoxicity of Xiangju cold granules and its protective effects on cells infected with influenza A viruses(H1N1,H3N2,oseltamivir-resistant H1N1)and other respiratory viruses(RSV,Sendai virus,and Adenoviruses)were detected in vitro with a CCK8 kit.Meanwhile,the effects of Xiangju cold granules on cytopathogenic effect(CPE)induced by virus infection were also observed.A mouse model of H1N1 virus nasal drip infection was established to evaluate the survival indexes of mice after Xiangju cold granules administration.The release of inflammatory factors and the activation of inflammatory signaling pathways in virus-infected mice after drug administration were analyzed by HE staining,ELISA,Western blot and immunohistochemistry.Results showed that the median cytotoxic concentration(CC50)of Xiangju cold granules was higher than its effective inhibitory concentration against these 6 tested virus strains,suggesting a good safety.It showed inhibitory activity against MDCK cytopathies caused by influenza A virus and Sendai virus,especially against oseltamivir-resistant H1N1.In vivo,Xiangju cold granules could significantly reduce the mortality of mice infected with H1N1,prolong animal survival time,reduce lung index,and relieve pulmonary inflammatory cells infiltration.Moreover,the release of lung inflammatory factors and the activation of NLRP3-caspase1-IL-1β protein were significantly decreased.In this study,cell models infected with 6 strains of viruses and mouse model infected by H1N were respectively established,demonstrating that Xiangju cold granules has good antiviral activity in vivo and in vitro,and can suppress the inflammatoryresponse caused by virus infection through blocking the NLRP3-caspase1-IL-1β pathway.
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