目的:探讨3种治法代表方剂对呼吸道合胞病毒(RSV)诱导引起肺炎的小鼠血清IL-6、IL-8及肺组织Toll样受体4(TLR4)、细胞核转录因子(NF—xB)的影响,揭示其治疗机制。方法:将Wister小鼠随机分为正常组、模型组、利巴韦林组、银翘散组、玉屏风散组、银屏散组。于感染前3d,玉屏风散组、银屏散组连续给予玉屏风散(7.5g·kg·d。)3d;除正常组以外,50μL 10LD50病毒液滴鼻建立呼吸道合胞病毒感染的小鼠RSV肺炎模型。感染2h后,银翘散组、银屏散组继续给予银翘散治疗(10g·kg·d-1),连续给药3d;采用ELISA法检测各组小鼠血清中IL-6、IL-8含量,免疫组化法检测分析肺组织TLR4、NF—κBP65蛋白的表达。结果:与正常组比较,RSV感染诱导小鼠肺组织中TLR4蛋白表达上调,增加NF—κB的活化(P〈0.05),使血清中IL-6、IL-8水平升高(P〈0.05)。银翘散组、玉屏风散组、双表法组测得IL-6、IL-8含量以及TL114、NF—κBP65表达相较于模型组显著减弱(P〈0.05)。与利巴韦林组比较,双表法组减弱尤为突出(P〈0.05)。结论:3种治法对于治疗RSV肺炎均有显著疗效,尤其以双表法疗效更佳。其作用机制可能与抑制炎症因子IL-6、IL-8分泌及阻断TLR4/NF-κB信号通路有关。
Objective : To investigate the influence of the representative prescriptions of three methods on expression of inflam- matory cytokines and toll - like receptor 4 ( TLR4 ) and nuclear factor KB ( NF -κB) in lung tissue of pneumonia mice models and to reveal the mechanism of the disease. Methods:48 Wister mice were randomly divided into 6 groups: normal group, model group, ribavirin group, Yinqiao Powder group, Yupingfeng group, Yinpin Powder group. Before 3 days of virus infection, Yupingfeng group and Yinpin Powder group were intragastrically given Yupingfeng at a dose of 7.5 g·kg- 1 . d - 1 by day 3. Except the normal group ,50 μL 10 LD50 RSV virus was intranasally inoculated to mice. 1 h after infection, mice in control and model groups were in- tragastrically given and mice in Yinqiao Powder and Yinping Powder group was intragastrically given Yinqiao Powder at a dose of 10 g·kg-1· d -1 by day 3. ELISA was used to the measure concentrations of IL -6 and IL - 8 in serum. Immunohistochemistry staining was used to detect the expressions of TLR4 and NF - κB P65 protein in lung tissue. Results : Compared with the normal group, respiratory syncytial virus could up - regulate the expression of TLR4 protein and increase the activation of NF - κB P65 in lnng tissue(P 〈0. 05) ,and enhance the levels of IL-6 and IL-8 in mice serum. The levels of IL-2 and IL-4 were increased in Yinqiao Powder group, Yupingfeng group and Yinpin Powder group, compared with those of the model control group, there was a significant difference (P 〈 0. 05 ). Compared with the ribavirin group, the expressions of those in Yinpin Powder group were weak- ened significantly. The expressions of those in Yinqiao Powder group were also lower than that of the ribavirin intragastric group( P 〈 0. 05 ). The decrease in Yupingfeng Powder group was not significant. Conclusion : The three methods have a very significant effect on RSV pneumonia, especially the Shuangbiao method group( P 〈 0. 05 ). The mechanism