金欣口服液含药血清对RSV感染RAW264.7细胞的RIG-I信号转导通路的影响  

Regulation effect of drug serum of Jinxin Oral Liquid on RIG-I signal transduction pathway in RAW264.7 cells infected with RSV

作　　者：陈争光[1] 汪受传[1] 魏肖云[1] 徐建亚[1] 

机构地区：[1]南京中医药大学中医儿科研究所,南京210029

出　　处：《中华中医药杂志》2014年第5期1338-1343,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(N o.81072840);全国高等学校博士学科点专项科研基金(No.20103237110003)~~

摘　　要：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。Objectve：To study the regulation effect of drug serum of Jinxin Oral Liquid on RIG- I signal transduction pathway in RAW264.7 ceils infected with RSV, and to explore its antiviral mechanism.Methods：RAW264.7 cells were infected with RSV, and were administered with the drug serum of Jin Xin Oral Liquid.After 12 hours, the cells were collected to detect the level of RIG- I mRNA, IPS- I mRNA, IRF3 mRNA and IFN-γ） mRNA with Real time RT-PCR and to measure the level of the protein of RIG- I, IPS- I, IRF3 and P-IRF3 by Western Blot, and the supernatant were collected to detect the level of IFN-γ） with ELISA assay.Results： The high expression of the mRNA of RIG- I, IPS- I, IRF3 and IFN-l） which was induced by RSV were down regulated significantly by the drug serum of Jinxin Oral Liquid （P〈0.01）, and the down-regulation effect of the drug serum of Jinxin Oral Liquid on IRF3 was lower than the Ribavirin （P〈0.05）. The high expression of the protein of RIG- I , IPS- I and P-IRF3 which was induced by RSV were down regulated significantly by the drug serum of Jinxin Oral Liquid （P〈0.01）, and the down-regulation effect on IPS-I and P-IRF3 was lower than the Ribavirin （P〈0.01, P〈0.05）, but the down-regulation effect on RIG- I was higher than the Ribavirin （P〈0.05）. The drug serum of Jinxin Oral Liquid had no effect on the expression of the protein of IRF3. The high expression of IFN-I） which was induced by RSV were down regulated significantly by the drug serum of Jinxin Oral Liquid （P〈0.01）, and the down-regulation effect of the drug serum of Jinxin Oral Liquid on IFN--β） was lower than the Ribavirin （P〈0.05）. Conclusion： The activation of RIG- I signaling transductlon pathway is induced by RSV to promote the expression of IFN-I）. The drug serum of Jin Xin Oral Liquid can inhibit the RIG- I signaling transduction pathway to control theproper expression of IFN--β.

关 键 词：金欣口服液含药血清 呼吸道合胞病毒 小鼠肺泡单核细胞 RIG-I信号转导通路 

分 类 号：R285.5[医药卫生—中药学]