芪丹复感颗粒对呼吸道病毒感染小鼠TLRs信号通路的影响

Effect of Qidan Fugan Granules on TLRs Signal Pathway of Mice Infected with Respiratory Virus

关键词:  流行性感冒  流感病毒  芪丹复感颗粒  Toll样受体

Key words:  influenza  influenza virus  Qidan Fugan granules  Toll-like receptors

摘要:

目的 探究芪丹复感颗粒对呼吸道病毒感染小鼠肺组织的保护作用及对TLRs信号通路的影响。方法 120只小鼠随机分为对照组，模型组，阳性药组（利巴韦林100 mg·kg^-1），芪丹复感颗粒低、中、高剂量组（芪丹复感颗粒1，2，4 g·kg^-1），每组20只，其中12只用于观察死亡情况。除对照组外，其余各组小鼠均通过鼻饲流感病毒PR8建立流感病毒感染模型。通过HE染色观察肺组织形态，ELISA检测小鼠血清和肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和白细胞介素-6（IL-6）的浓度。分别使用Western blot和RT-qPCR检测Toll样受体4/7（TLR4/7）、髓样分化因子（MyD88）、核因子κB（NF-κB）蛋白表达和mRNA水平。结果 建模第7天，模型组小鼠全部死亡，死亡率为100.0%。在建模后第14天阳性药组死亡3只（25.0%），芪丹复感颗粒低、中、高剂量组分别死亡9只（75.0%）、6只（50.0%）和5只（41.7%）。造模第5天，模型组，阳性药组，低、中、高剂量组的血清和BALF中的TNF-α、IL-6、IL-1β水平以及肺组织中TLR4/7、MyD88、NF-κB蛋白表达和mRNA水平均显著高于对照组（P<0.05），阳性药组，低、中、高剂量组均显著低于模型组（P<0.05），中、高剂量组显著低于低剂量组（P<0.05），而中、高剂量组之间无显著差异。结论 芪丹复感颗粒可以减轻流感病毒造成的肺损伤，可能是通过抑制TLR4/7-MyD88-NF-κB通路的表达抑制炎症因子的释放，从而降低肺损伤。

Abstract:

OBJECTIVE To explore the protective effect of Qidan Fugan granules on lung tissue of mice infected with respiratory virus and its effect on TLRs signaling pathway. METHODS One hundred and twenty mice were randomly divided to control group, model group, positive drug group(ribavirin 100 mg·kg^-1), low, medium and high dose groups(Qidan Fugan granules 1, 2, 4 g·kg^-1), 20 mice in each group, 12 of them were used to observe deaths. Except the control group, the other groups of mice established a model of influenza virus infection by nasal feeding PR8. The morphology of lung tissue was detected by HE staining. The concentration of TNF-α, IL-1β, IL-6 in serum and bronchoalveolar lavage fluid(BALF) were detected by ELISA. Western blot and RT-qPCR were used to detect TLR4/7, MyD88, NF-κB protein and mRNA levels. RESULTS On the 7th day, all mice in the model group died, with a mortality rate of 100.0%. On the 14th day after the establishment of the model, 3(25.0%) died in the positive drug group, 9(75.0%), 6(50.0%) and 5(41.7%) died respectively in the low, medium and high dose groups. On the 5th day, in the model group, positive drug group, low, medium and high dose groups the levels of TNF-α, IL-6, IL-1β in serum and BALF, and the expression of TLR4/7, MyD88, NF-κB protein, the levels of mRNA in lung tissue were significantly higher than the control group(P<0.05). The low, medium and high dose group were significantly lower than the model group(P<0.05), and the middle and high dose groups were significantly lower than the low dose group(P<0.05). There was no significant difference between the medium and high dose groups. CONCLUSION Qidan Fugan granules can alleviate lung injury caused by influenza virus. It may be to inhibit the release of inflammatory factors by inhibiting the expression of TLR4/7-MyD88-NF-κB pathway, thereby reducing lung injury.