香烟烟雾对小鼠肺泡巨噬细胞RIG-I样受体功能影响的研究

Effect of cigarette smoke extract on RIG-I-like receptor in mouse alveolar macrophages

目的探讨吸烟对肺泡巨噬细胞RIG-I样受体介导的信号传导功能。方法制备香烟烟雾提取物（cigarettesmokeextract，CSE），用病毒模拟物Poly（I：C）及不同浓度CSE单独或者联合刺激小鼠肺泡巨噬细胞（MHs）。收集细胞并抽提RNA，用实时荧光定量PCR检测RIG-I、MDA-5、干扰素8（IFNB）、IL-1及IL-6mRNA表达水平。结果单独的CSE刺激能引起IL-1、IL-6mRNA等炎症细胞因子的水平升高，而对RIC-I、MDA-5及IFN-0mRNA几乎无影响；CSE能够抑制Poly（I：C）刺激引起的RIG-I、MDA5受体及分子IFN—β、IL-1、IL-6增高现象，尤其对IFN—βmRNA的抑制最为明显，并且CSE的这种抑制作用与其浓度呈正相关。结论单独CSE刺激MH—S细胞可以引起炎症因子的表达水平升高，Poly（I：c）刺激引起的RIG-I样受体抗病毒信号通路中相关受体、IFN及炎症因子表达水平增高现象可以被CSE抑制，巨噬细胞对病毒防御功能的减低可能与吸烟抑制RIG-I样受体信号通路功能有关。

Objective To investigate the effect of cigarette smoke extract （CSE） on mouse alveolar macrophage after stimulation of RIG-I-like receptor （RLR）. Methods CSE was prepared,and then MH-S was treated with Poly （I：C） or CSE or the two together. And RNA was isolated from the indicated mouse alveoar macrophages. Realtime PCR was performed to measured the expressions of RIG-I, MDA-5, interferon-β （IFN-β） ,interleukin-1 （IL-1） ,and IL-6 mRNA. Results CSE increased IL-1 and IL-6 mRNA expressions, while it had no effect on RIG-I, MDA-5, and IFN β mRNA expressions. Poly （I： C） stimulation not only increased RIG-I and MDA-5 receptor levels, but also elevated IFN 13, IL 1, and IL-6 mRNA expressions. While this phenomenon could be attenuated by CSE, this attenuation was very significant for IFN-β. Conclusions CSE can elevate the expressions of inflammatory cytokines,it can also inhibit the expressions of related receptor, interferon, and inflammatory cytokines in the RLR antiviral signaling pathways after Poly （I：C） stimulation. The reduction of defense to virus in alveolar macrophages may be related to the influence of smoking on the RLR signaling pathways.