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目的通过检测鼠流感的肺病理变化及脾细胞Th1/Th2免疫因子及气管粘膜TLR2/4 mRNA的表达变化,初步探讨冰香散挥发油制剂对流感病毒的防治作用及其免疫调节机制。方法选用BALB/c小鼠,随机分为正常组、病毒对照组、利巴韦林组、冰香散挥发油制剂滴鼻组、雾化组,采用不同的给药方式预防性给药7d,第8天以15LD50攻毒,再持续给药5天后取材。肺组织经HE染色后观察其病变程度;脾脏研碎制得脾细胞悬浮液,以RT-PCR法检测细胞因子IFN-γ、IL-4、IL-5的分泌情况;RT-PCR检测气管粘膜上TLR2/4基因表达的水平。结果小鼠经H1N1感染后肺部出现严重的炎性细胞浸润,与正常组相比TLR2/4 mRNA、IFN-γmRNA表达水平上升(P<0.01),IL-4、L-5 mRNA表达水平下降(P<0.01)。与病毒对照组相比利巴韦林组可显著增加IL-4 mRNA的表达(P<0.01),降低IFN-γmRNA表达(P<0.05);冰香散挥发油制剂滴鼻、雾化组能显著增加IL-4 mRNA、IL-5 mRNA的表达水平(P<0.01或P<0.05),但在降低IFN-γmRNA的表达上冰香散挥发油制剂滴鼻组的效果比较显著(P<0.05)。各给药组都有降低TLR2/4基因表达的趋势,但只有冰香散挥发油制剂滴鼻组的作用比较显著(P<0.01)。结论冰香散挥发油制剂对流感病毒性肺炎病变有一定的防治作用,调控炎性细胞因子及TLR2/4基因的表达水平可能是其抗流感所致肺炎的重要作用机制之一。
OBJECTIVE: To investigate the protective effect of Bingxian San volatile oil on influenza virus and its immunoregulatory mechanism by detecting the pathological changes of lungs in mice and the expression of Th1 / Th2 immune factors and TLR2 / 4 mRNA in the tracheal mucosa. Methods BALB / c mice were randomly divided into normal group, virus control group, ribavirin group and volatile oil preparation of Rhizoma Pinelliae Preparata nasal spray group and nebulization group. Different administrations were given for prophylactic administration for 7 days. 8 days to 15LD50 challenge, and then continue to administer 5 days after drawing. The degree of lesion was observed by HE staining. The spleen cells were pulverized to obtain the spleen cell suspension. The secretion of cytokines IFN-γ, IL-4 and IL-5 were detected by RT-PCR. On the level of TLR2 / 4 gene expression. Results Serum of inflammatory cells infiltrated the lungs of mice infected with H1N1. Compared with the normal group, the expression of TLR2 / 4 mRNA and IFN-γ mRNA increased (P <0.01), while the expression of IL-4 and L-5 mRNA decreased (P <0.01). Compared with the virus control group, ribavirin group could significantly increase the expression of IL-4 mRNA (P <0.01) and reduce the expression of IFN-γmRNA (P <0.05) (P <0.01 or P <0.05), but decreased the expression of IFN-γ mRNA significantly (P <0.05). Each administration group had the tendency to reduce the expression of TLR2 / 4 gene, but only the effect of the volatile oil of Bingxiang San on nasal drops group was significant (P <0.01). Conclusion Bingxian San volatile oil preparation has some preventive and therapeutic effects on the pathogenesis of influenza virus pneumonia. The regulation of the expression of inflammatory cytokines and TLR2 / 4 may be one of the important mechanisms of anti-influenza pneumonia.