目的 研究清热消炎宁抗冠状病毒的作用，为评价其防治冠状病毒感染提供实验依据。方法 雌雄各半的96只BALB/c小鼠随机分为对照组、模型组、连花清瘟胶囊(0.546 g/kg)组和清热消炎宁片低、中、高剂量(8.72、17.44、34.89 g/kg)组，每组16只。采用ip环磷酰胺联合HCoV-229E冠状病毒感染BALB/c小鼠，建立冠状病毒感染模型，通过小鼠体质量、肺指数、病毒载量、血凝滴度、肺组织病理改变来评价清热消炎宁的治疗作用；通过ELISA检测肺泡灌洗液中白细胞介素-1β(interleukin-1β，IL-1β)、IL-4、肿瘤坏死因子-α(tumor necrosis factor-α，TNF-α)、γ干扰素(interferon-γ，IFN-γ)和血管细胞黏附分子-1(vascular cell adhesion molecule-1，VCAM-1)水平；流式细胞术检测肺组织巨噬细胞、淋巴细胞(CD3⁺、CD4⁺)及NK细胞比例；Western blotting检测肺组织Toll样受体4(Toll-like receptor 4，TLR4)、髓样分化因子88(myeloid differentiation factor 88，MYD88)、抑制因子κB激酶-β(inhibitor kappa B kinase-β，IKK-β)、抑制因子κB(inhibitor kappa B，IκB)和p-IκB蛋白表达。结果 与模型组比较，清热消炎宁明显增加病毒感染的小鼠体质量(P＜0.05、0.01)，降低肺指数和血凝效价(P＜0.01)，改善肺脏病变(P＜0.05)，且能显著抑制病毒mRNA表达(P＜0.01)；降低肺泡灌洗液中TNF-α、IL-1β和VCAM-1水平(P＜0.05、0.01)，提高IFN-γ水平(P＜0.05)；明显降低巨噬细胞比例(P＜0.05、0.01)，提高淋巴细胞CD3⁺、CD4⁺和NK细胞比例(P＜0.01)；显著下调肺组织MYD88、TLR4、IκB、IKK-β蛋白表达水平(P＜0.05、0.01)。结论 清热消炎宁片可以抑制冠状病毒体内复制，减轻炎症反应，保护肺脏组织，具有明显的体内抗冠状病毒的药效作用，其作用机制可能与调控TLR4/MyD88/IKK/IκB信号通路和提高免疫有关。

Objective To study the anti-coronavirus effect of Qingre Xiaoyanning Tablet (清热消炎宁片), and provide experimental basis for evaluating its prevention and treatment of coronavirus infection. Methods A total of 96 BALB/c mice with half male and half female were randomly divided into control group, model group, Lianhua Qingwen Capsules (连花清瘟胶囊, 0.546 g/kg) group and Qingre Xiaoyanning Tablet (8.72, 17.44, 34.89 g/kg) groups with 16 mice in each group. BALB/c mice were infected with ip cyclophosphamide combined with HCoV-229E coronavirus to establish a model of coronavirus infection. The therapeutic effect of Qingre Xiaoyanning Tablet was evaluated by body weight, lung index, viral load, hemagglutination titer and pathological changes in lung tissue of mice; Levels of interleukin-1β (IL-1β), IL-4, tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and vascular cell adhesion molecule-1 (VCAM-1) in alveolar lavage fluid were detected by ELISA; The proportion of macrophages, lymphocytes (CD3⁺, CD4⁺) and NK cells in lung tissue was detected by flow cytometry; Western blotting was used to detect Toll like receptor 4 (TLR4), myeloid differentiation factor 88 (MYD88), inhibitor kappa B kinase-β (IKK-β), inhibitor kappa B (IκB) and p-IκB protein expressions in lung tissue. Results Compared with model group, Qingre Xiaoyanning Tablet significantly increased the body weight of virus infected mice (P＜0.05, 0.01), decreased lung index and hemagglutination titer (P＜0.01), improved lung disease (P＜0.05), and significantly inhibited viral mRNA expression (P＜0.01); TNF-α, IL-1 β and VCAM-1 levels in alveolar lavage fluid were decreased (P＜0.05, 0.01), IFN-γ level was increased (P＜0.05); The percentage of macrophages was significantly decreased (P＜0.05, 0.01), percentage of CD3⁺, CD4⁺ lymphocytes and NK cells was increased (P＜0.01); MYD88, TLR4, IκB and IKK-β protein expressions in lung tissue were significantly down regulated (P＜0.05, 0.01). Conclusion Qingre Xiaoyanning Tablet can inhibit the replication of coronavirus in vivo, reduce inflammatory reaction, protect lung tissue, and has obvious anti-coronavirus effect in vivo. Its mechanism may be related to the regulation of TLR4/MyD88/IKK/IκB signal pathway and improving immunity.

R285.5

湖南省重点研发计划(2020DK2003)；自然科学基金-科药联合基金资助项目(2022JJ80030)