Objective: To investigate the expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei(SFXF) and Jiebiaoqingli (JBQL) Chinese herbal anti-virus formulas.
Methods: Mice were anesthetized and then infected intranasally by dropping 0.05 mL of influenza virus suspension (4×LD50;) except normal group. Mice were divided randomly into nine groups: normal group, model group (virus only), control group [11.375 g/(kg.d)Oseltamivir], low-dose SFXF [0.94 g/(kg.d)], medium-dose SFXF [1.88 g/(kg.d)], high-dose SFXF [3.76 g/(kg.d)], low-dose JBQL [1.09 g/(kg.d)], medium-dose JBQL [2.18 g/(kg.d)] and high-dose JBQL [4.36 g/(kg.d)]. Oseltamivir group, SFXF groups and JBQL groups were administered to mice by oral gavage in equal dose of 0.2 mL daily for 4 consecutive days, while the rest of the groups received water only. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Select differentially expressed genes of cytokines involved in inflammation. Some candidate genes, such as IL-1β, IL-8, IL-10, RANTES and ICAM-1 were verified by qRT-PCR. To confirm the genes expression data from the microarray involved in inflammation in response to virus infection and treatment, we used qPCR to verify mRNA relative expressions of IL-1β, IL-8, IL-10, RANTES and ICAM-1. The expression of IL-1β protein in lung tissues was verified by Western blotting.
Results: IL-1β, CXCR2, CCL5, IL-10, IL-6, IL-18, TGF-β1 and CCL2 were up-regulated in model group. Gene expressions of IL-1β, CXCR2, CCL5, IL-10 and IL-6 were significantly down-regulated by all therapeutic groups. SFXF in medium-dose and low-dose down-regulated gene expressions of IL-18, TGF-β1, CCL2 and CCL5. IL-18 and CCL5 was down-regulated by both low-dose and medium-dose JBQL. qRT-PCR and western blot experiments showed that two formulas in medium-dose can down-regulate mRNA and protein expression of IL-1β (P<0.01). Both SFXF and JBQL in medium-dose significantly decreased the IL-8, RANTES, ICAM-1 and IL-10 mRNA expression (P<0.05 or P<0.01), compared with the model group. As expected, qRT-PCR data were in good agreement with the microarray assay.
Conclusion: The two anti-viral formulas may inhibit inflammatory immunopathogenesis, and may have the actions of protection the lung tissue from influenza-induced injury.