银花平感颗粒和三拗汤体内抗甲型流感病毒的免疫机制研究

Immunologic mechanisms of Yinhua Pinggan granule and San-ao decoction against influenza virus in vivo

为探讨银花平感颗粒和三拗汤体内对流感病毒肺炎小鼠免疫调节机制的影响及不同方剂配伍对流感病毒的药效异l司。米用滴鼻感染建立小鼠甲型流感病毒（H1N1）肺炎模型，随机分为正常对照组、病毒模型组、达菲组、银花平感颗粒组、三拗汤组，灌胃给予相应药物5d后，处死动物。酶联免疫吸附法（ELISA）检测流感病毒感染小鼠血清中肿瘤坏死因子-α（TNF-α）、臼细胞介素石（IL一6）和干扰素-γ（IFN-γ）的含量，荧光定量PCR法检测肺组织中T0u样受体3／7（TLR3／7）、髓样分化因子88（MyD88）、c-Jun氨基末端激酶（JNK）、丝裂原活化蛋白激酶（p38MAPK）、转录因子-KB（NF-KBp65）基因的表达，免疫组化法检On．0JNK，p38MAPK，NF．KBp65蛋白的表达。结果显示，银花平感颗粒和三拗汤均显著降低肺组织血清中TNF-α和IL-6水平及升高IFN-γ水平，显著下调流感病毒感染小鼠肺组织中TLR3／7，MyD88，JNK，p38MAPK，NF-KBp65基因的表达水平，显著降低肺组织中JNK，p38MAPK，NF-KBp65蛋白的表达水平，且银花平感颗粒对其调控效果优于三拗汤。结果表明，银花平感颗粒和三拗汤对流感病毒肺炎小鼠有一定的治疗作用，其抗流感病毒的作用机制可能是通过调节流感病毒感染小鼠的免疫功能和调控TLR3／7信号通路多个环节的基因和蛋白表达相互作用的结果，兼有清热解毒与宣肺解表疏邪之法的辛温与辛凉合用药银花平感颗粒比辛温单用药三拗汤配伍更全面，疗效更好。

This paper aimed to investigate the effect of Yinhua Pinggan granule and San-ao decoction on the immunologic mecha- nisms of influenza viral pneumonia mice in vivo, in order to study the activity of the combined administration of different formulas on in- fluenza A/H1 N1 virus. The model of pneumonia was established in mice through nasal dropping influenza virus, and then divided ran- domly into five groups： normal control group, influenza virus model group, oseltamivir control group, Yinhua Pinggan granule group, and San-ao decoction group. The animals were put to death at the 5th day after garage administration with the corresponding drugs. The contents in mice serum of TNF-α, IL-6 and IFN-γ were respectively measured by ELISA. The mRNA expressions of TLR3/7, MyD88, JNK, p38MAPK and NF-KB p65 in lung tissues were respectively detected by RT-PCR. The protein expressions of JNK, p38MAPK and NF-KB p65 in lung tissues were determined by immunohistochemical analysis, respectively. According to the results, Yinhua Ping- gan granule and San-ao decoction could significantly decrease the levels of TNF-α and IL-6, increase the level of IFN-T in mice serum of lung tissues, significantly reduce the gene expressions of TLR3/7, MyDS8, JNK, p38MAPK and NF-KB p65 in influenza virus-in- fected mice lung tissues, and significantly reduce the protein expressions of JNK, p38MAPK and NF-KB p65 in lung tissues. Further-more, the regulatory effect of Yinhua Pinggan granule was superior to that of San-ao decoction. In conclusion, Yinhua Pingan granule and San-ao decoction have the therapeutic effect on pneumonia mice infected by H1N1 virus in vivo. The anti-influenza mechanisms of Yinhua Pinggan granule and San-ao decoction may be the results of interactions by regulating the immunologic function of influenza vi- rus-infected mice and TLR3,/7 signaling pathway with multiple links of the gene and protein expressions. Moreover, the combined ad- ministration of warm-natured and cold-natured Yinhua Pinggan granule with the effects of detoxification and exhalation has a better effect than the single administration of warm-natured San-ao decoction.