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目的:研究金欣口服液含药血清对呼吸道合胞病毒(RSV)感染小鼠肺泡单核细胞(RAW264.7)中维甲酸诱导基因Ⅰ(RIG-Ⅰ)信号转导通路的影响,探讨其可能的抗病毒机制。方法:RSV感染RAW264.7细胞,采用金欣口服液含药血清进行干预,12h后收集细胞及其上清液,Real-time RT-PCR检测RAW264.7细胞RIG-Ⅰ、IPS-Ⅰ、IRF3和IFN-βmRNA表达情况;Western Blot法检测RIG-Ⅰ、IPS-Ⅰ、IRF3和P-IRF3;ELISA法检测细胞上清液中IFN-β含量。结果:金欣口服液含药血清能够显著下调RSV诱导的RIG-Ⅰ、IPS-Ⅰ、IRF3和IFN-βmRNA的高表达(P<0.01),但对IRF3的下调作用低于利巴韦林组(P<0.05)。金欣口服液含药血清能够显著下调RSV诱导的RIG-Ⅰ、IPS-Ⅰ和P-IRF3蛋白的高表达(P<0.01),但对IPS-Ⅰ和P-IRF3的下调作用低于利巴韦林组(P<0.01,P<0.05),而对RIG-Ⅰ的下调作用高于利巴韦林组(P<0.05)。金欣口服液含药血清对IRF3蛋白无明显调控作用。金欣口服液含药血清能够显著下调RSV诱导的IFN-β的高表达(P<0.01),但对IFN-β的下调作用低于利巴韦林组(P<0.05)。结论:RSV能够诱导RIG-Ⅰ信号转导通路的激活从而促进IFN-β的表达,而金欣口服液含药血清通过抑制RIG-Ⅰ信号转导通路使IFN-β的适度表达。
Objective: To investigate the effect of Jinxin oral liquid-containing serum on the signal transduction pathway of retinoic acid-induced gene Ⅰ (RIG-Ⅰ) in alveolar monocytes (RAW264.7) infected with respiratory syncytial virus (RSV) Possible antiviral mechanisms. Methods: RAW264.7 cells were infected with RSV, and the cells were infected with Jinxin oral liquid. After 12 hours, the cells and their supernatants were collected. Real-time RT-PCR was used to detect RIG-Ⅰ, IPS-Ⅰ, IRF3 And IFN-βmRNA expression; Western Blot detection of RIG-Ⅰ, IPS-Ⅰ, IRF3 and P-IRF3; ELISA assay of cell supernatant IFN-β content. Results: The serum containing Jinxin oral liquid could significantly down-regulate the expression of RIG-Ⅰ, IPS-Ⅰ, IRF3 and IFN-βmRNA induced by RSV (P <0.01), but downregulated IRF3 was lower than that of ribavirin (P <0.05). Jinxin oral liquid-containing serum could significantly downregulate RSV-induced high expression of RIG-I, IPS-I and P-IRF3 protein (P <0.01), but downregulation of IPS-I and P-IRF3 (P <0.01, P <0.05), while the effect of RIG-Ⅰ was lower than that of ribavirin group (P <0.05). Jinxin oral liquid containing serum IRF3 protein no significant regulatory role. Jinxin oral liquid containing serum could significantly down-regulate the expression of IFN-β induced by RSV (P <0.01), but lower the effect of IFN-β than ribavirin group (P <0.05). Conclusion: RSV can induce the activation of RIG-Ⅰ signal pathway and promote the expression of IFN-β, while Jinxin oral liquid containing serum can moderate the expression of IFN-β by inhibiting RIG-Ⅰ signal transduction pathway.