金振口服液抑制小鼠流感病毒性肺炎的药效及机制研究

Effect and mechanism of Jinzhen Oral Liquid on influenza virus pneumonia in mice

目的 研究金振口服液抑制小鼠流感病毒性肺炎的药效及作用机制。方法 将ICR小鼠随机分为对照组、模型组,金振口服液高、中、低剂量(4.4、2.2、1.1 g/kg)组和磷酸奥司他韦胶囊(0.05 g/kg)组。除对照组外,其余各组小鼠滴鼻感染甲型H1N1流感病毒构建小鼠病毒性肺炎模型,测定各组小鼠存活时间和肺指数;苏木素–伊红(HE)染色法观察肺组织病理变化;流式细胞术检测外周血CD3^(+)、CD4^(+)、CD8^(+)T淋巴细胞水平;ELISA法检测血清及肺泡灌洗液中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)及白细胞介素-10(IL-10)水平;Western blotting法检测肺组织Toll样受体3(TLR3)、β干扰素TIR结构域衔接蛋白(TRIF)和核转录因子-κB(NF-κB)蛋白表达。结果 与模型组相比,金振口服液给药组和磷酸奥司他韦胶囊组能显著延长小鼠存活时间,降低肺指数(P<0.05、0.01);减轻肺组织病理损伤;显著上调外周血CD3^(+)、CD4^(+)T淋巴细胞水平及CD4^(+)/CD8^(+)值,下调CD8^(+)T淋巴细胞水平(P<0.05、0.01);明显降低血清及肺泡灌洗液中TNF-α、IL-1β和IL-6水平,升高IL-10水平(P<0.05、0.01);显著抑制肺组织TLR3、TRIF及NF-κB蛋白表达(P<0.05、0.01)。结论 金振口服液能够延长病毒性肺炎小鼠存活时间、降低肺指数,减轻肺部炎性损伤,其抑制小鼠流感病毒性肺炎的机制可能与抑制TLR3/TRIF信号传导、调节抗炎–促炎失衡和改善T细胞免疫有关。

Objective To study the efficacy and mechanism of Jinzhen Oral Liquid in inhibiting influenza virus pneumonia in mice.Methods ICR mice were randomly divided into control group, model group, Jinzhen Oral Liquid high, medium, and low dose(4.4,2.2, 1.1 g/kg) groups, and Oseltamivir Phosphate Capsules(0.05 g/kg) group. Except for the control group, mice in other groups were intranasally infected with influenza A(H1N1) virus to build a mouse viral pneumonia model. Survival time and lung index of mice in each group were measured. The pathological changes of lung tissue were observed by hematoxylin-eosin(HE) staining. The levels of CD3^(+), CD4^(+), CD8^(+)T lymphocytes in peripheral blood were detected by flow cytometry. Detection of TNF-α, IL-1β, IL-6, and IL-10levels in serum and alveolar lavage fluid were carried out by ELISA. Western blotting method was used to detect TLR3, TRIF, and NF-κB protein expression. Results Compared with the model group, the survival time of mice in Jinzhen Oral Liquid group and Oseltamivir Phosphate Capsules group were significantly prolonged, and the lung index was reduced(P < 0.05, 0.01). The pathological results also showed alleviated damage of lung tissue in these two groups. The level of CD3^(+), CD4^(+)T lymphocytes and the ratio of CD4^(+)/CD8^(+)in peripheral blood were largely increased, and the level of CD8^(+)T lymphocytes was greatly decreased(P < 0.05, 0.01).TNF-α, IL-1β and IL-6 content in serum and alveolar lavage fluid were reduced, while IL-10 content was increased(P < 0.05, 0.01).The expression of TLR3, TRIF, and NF-κB were significantly inhibited in lung tissue(P < 0.05, 0.01). Conclusion Jinzhen Oral Liquid can prolong the survival time of mice with viral pneumonia, reduce lung index, and alleviate lung inflammatory injury. Its mechanism of inhibiting influenza viral pneumonia in mice may be related to the inhibition of TLR3/TRIF signal transduction,regulation of anti-inflammatory and pro-inflammatory imbalance, and improvement of T cell immunity.