清热消炎宁抗甲型流感H3N2病毒的药效及作用机制研究  

Study on the efficacy and mechanism of Qingre xiaoyanning against influenza A H3N2 virus

作　　者：周莎莎 程雪清 彭冬冬 王小青 扶丽君 肖文喜 张国民[1] ZHOU Shasha;CHENG Xueqing;PENG Dongdong;WANG Xiaoqing;FU Lijun;XIAOWenxi;ZHANG Guomin(Hunan University of Chinese Medicine,School of Integrated Chinese and Western Medicine,Changsha 410208,Hunan,China;Hinye Pharmaceutical Co.,Ltd.,Medicine Research Institute,Changsha 410331,Hunan,China;Hunan Key Laboratory of Pharmacodynamics and Safety Evaluation of New Drugs,Drug Evaluation Department,Changsha 410331,Hunan,China;Human Prima Drug Research Center Co.,Ltd.,Drug Evaluation Department,Changsha 410331,Hunan,China)

机构地区：[1]湖南中医药大学,中西医结合学院,湖南长沙410208 [2]天地恒一制药股份有限公司,药物研究院,湖南长沙410331 [3]新药药效与安全性评价湖南省重点实验室,药物评价部,湖南长沙410331 [4]湖南普瑞玛药物研究中心有限公司,药物评价部,湖南长沙410331

出　　处：《中国临床药理学与治疗学》2025年第3期347-354,共8页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：湖南省重点研发计划项目(2023SK2080)。

摘　　要：目的:研究清热消炎宁(QRXYN)体内抗病毒药效及作用机制,为其防治甲型流感提供实验依据。方法:构建感染甲型流感H3N2病毒小鼠模型,通过小鼠体质量变化、肺组织病理改变、血凝滴度、病毒载量评价QRXYN抗甲型流感病毒的药效作用;通过ELISA检测小鼠肺泡灌洗液中TNF-α、IL-1β、IL-4、IFN-γ、血管细胞黏附分子-1(VCAM-1)的水平;通过流式细胞术检测肺组织中巨噬细胞(F4/80)、辅助T淋巴细胞(CD4^(+)T淋巴细胞)及自然杀伤(NK)细胞的比例;通过免疫印迹法(WB)检测肺组织Toll样受体4(TLR4)、髓样分化因子88(MYD88)、抑制因子κB激酶-β(IKK-β)、核因子κB抑制因子α(IκBα)和磷酸化IκB激酶-α(p-IκBα)蛋白的表达。结果:与模型组比较,奥司他韦及清热消炎宁均能减轻小鼠肺组织病变程度,降低小鼠肺组织血凝滴度及病毒载量(P<0.01),降低肺泡灌洗液中TNF-α、IL-4、VCAM-1的水平(P<0.05,P<0.01),减少巨噬细胞比例(P<0.05,P<0.01),提高CD4^(+)T淋巴细胞及NK细胞比例(P<0.05,P<0.01);此外奥司他韦可降低小鼠肺脏MYD88蛋白表达(P<0.05),清热消炎宁可降低小鼠肺脏IKK-β与P-IκBα蛋白的表达(P<0.05)。结论:清热消炎宁具有良好的体内抗甲型流感病毒作用,其机制可能与调节免疫功能和NF-κB信号通路有关。AIM:To investigate the antiviral efficacy and mechanism of Qingre Xiaoyanning(QRXYN)in vivo,and provide experimental basis for their prevention and treatment of influenza A virus.METHODS:We constructed a mouse model infected with influenza A H3N2 virus.To evaluate the therapeutic effect of QRXYN on influenza A virus,we measured the body weight changes,pathological changes in lung tissue,hemagglutination titer,and viral load in mouse.To evaluate the possible mechanism of QRXYN's anti influenza A virus infection,we used the ELISA to measure the levels of TNF-α,IL-1β,IL-4,IFN-γ,and vascular cell adhesion molecule-1(VCAM-1)in mouse bronchoalveolar lavage fluid;used flow cytometry to assess the proportions of macrophages(F4/80),helper T lymphocytes(CD4^(+)T lymphocytes),and natural killer(NK)cells in lung tissue;and used Western blotting to detect the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MYD88),inhibitor of kappa B kinase-β(IKK-β),NF-kappa-B inhibitor alpha(IκBα),and phospho-IKB alpha(p-IκBα)in lung tissue.RESULTS:Compared to the model group,both Oseltamivir and QRXYN can alleviate the severity of lung tissue lesions in mice,decrease the blood coagulation titer and viral load of mouse lung tissue(P<0.01),lower the levels of TNF-α,IL-4,and VCAM-1 in bronchoalveolar lavage fluid(P<0.05,P<0.01),reduce the proportion of macrophages(P<0.05,P<0.01),and increase the proportion of CD4^(+)T lymphocytes and NK cells(P<0.05,P<0.01).Additionally,oseltamivir can reduce the expression of MYD88 protein in mouse lungs(P<0.05),while QRXYN can decrease the expression of IKK-βand P-IκBαproteins in mouse lungs(P<0.05).CONCLUSION:QRXYN have good in vivo antiviral effects against the influenza A virus,and their mechanism may be related to the regulation of the immunologic function and NF-κB signal pathway.

关 键 词：甲型流感 髓样分化因子88 奥司他韦 肺泡灌洗液 血凝滴度 IΚB激酶 T淋巴细胞 巨噬细胞 

分 类 号：R285.5[医药卫生—中药学]