基于microRNAs的石荠苧总黄酮抗流感病毒性肺炎作用机制研究

Mechanism of total flavonoids from Mosla scabra on resistance to influenza viral pneumonia based on micro RNAs

目的从微小核糖核酸(micro RNAs,mi RNAs)角度研究石荠苧总黄酮抗流感病毒性肺炎作用机制。方法设立对照组、模型组和石荠苧总黄酮组,将甲型流感病毒小鼠肺适应株A/PR/8/34经鼻滴入模型组和石荠苧总黄酮组小鼠,建立小鼠流感病毒感染性肺炎模型,观察比较石荠苧总黄酮对流感病毒感染小鼠肺指数和血清白细胞介素-6(IL-6)、γ干扰素(IFN-γ)水平的影响;采用高通量测序法和荧光定量PCR法,检测各组小鼠肺组织中mi RNAs的表达丰度差异;采用miranda、mirbase、targetscan 3个数据库预测差异mi RNAs的靶基因,并通过KEGG分析靶基因的相关功能;采用蛋白印迹法验证相关蛋白的表达。结果与模型组相比,石荠苧总黄酮可显著抑制流感病毒感染引起的小鼠肺指数和血清中细胞因子增加,调节异常表达的mi RNAs水平趋于正常;KEGG分析这些mi RNAs所调控的靶基因主要富集于JAK-STAT、TLR3等信号通路;Western blotting证实石荠苧总黄酮可调整感染小鼠异常表达的靶蛋白水平。结论石荠苧总黄酮可通过调控小鼠体内mi RNAs的表达发挥抗流感病毒性肺炎作用。

Objective To investigate the mechanism of flavonoids from Mosla scabra（FMS） on anti-influenza from the sight of micro RNAs. Methods Mice were divided into normal group, model（MC） group, and FMS group. Mice in MC and FMS groups were infected with influenza virus H1N1, then mice in the FMS group were treated with FMS. To observe the influence of mice in FMS group for the lung index and the levels of cytokines in serum. The difference expressing of mi RNAs in lung tissue of mice in each group were detected by high-flux sequencing and quantitative real-time PCR. Human m RNA database as target was used to predict the target genes of differentially expressed mi RNAs by miranda, mirbase, and targetscan analysis, while the target genes functions were considered by KEGG analyses. The related proteins of target genes were tested by Western blotting. Results FMS could significantly decrease the lung index and cytokines of infected mice and regulate the expression levels of abnormal mi RNAs tend to normal. We also found that mi RNAs are relevant to JAK-STAT and TLR3 signal pathways by KEGG. Western blotting confirmed that FMS could adjust the abnormal protein level of infected mice. Conclusion FMS obviously alleviates viral pneumonia via regulating mi RNA expression in mice.