参苏饮含药血清对小鼠巨噬细胞Toll样受体及其下游信号转导通路的影响

目的：研究参苏饮含药血清对脂多糖(LPS)和聚肌胞(POLY I∶C)刺激的小鼠巨噬细胞株RAW 264.7表达Toll样受体3(TLR3),Toll样受体4(TLR4)及其下游信号转导通路髓样分化蛋白(MyD88),肿瘤坏死因子受体相关因子-6(TRAF-6),Toll样受体相关分子(TRAM)和Toll样受体相关的干扰素活化子(TRIF) mRNA的影响。方法：用LPS(5 μg·mL^-1),POLY I∶C(50 μg·mL^-1)分别刺激RAW 264.7细胞株,同时给予参苏饮含药血清(11.2,6.7,4.0,2.4,1.5 g·kg-1·U^-1)进行干预,ELISA法测定细胞培养上清液中炎症因子肿瘤坏死因子-α(TNF-α)和干扰素-β (IFN-β)的含量。先计算参苏饮IC₅₀,然后再以IC₅₀(11,5,2.5,4.9 g·kg^-1·U^-1)做为药物剂量进行干预,荧光定量PCR方法测定TLR3,TLR4和下游通路相关指标mRNA的表达,分析评价参苏饮含药血清清热解毒药效学作用的现代药理学基础。结果：参苏饮含药血清对POLY I∶C刺激的TLR3及其下游通路的MyD88和TRAM有抑制作用,对LPS刺激的TLR4的病理性升高无抑制作用,但对TLR4下游通路TRAM和TRIF mRNA的表达有明显的抑制作用。以上综合作用引起炎症因子TNF-α和IFN-β的降低。结论：参苏饮清热解毒的药效学作用可能与抑制TLR3,MyD88,TRAM和TRIF等细胞因子的表达有关。

Effect of Shensuyin on the expression of toll-like receptors and the downstream signaling components on RAW 264.7 cells

OBJECTIVE: To investigate the influences of Shensu Yin to RAW 264.7 on the expression of TLR3, TLR4 and the factors of the downstream in RAW 264. 7 cells. METHOD: RAW 264.7 cell line was stimulated with Lipopolysaccharide and POLY I: C, respectively, and treated with the drug serum of Shensuyin simultaneously. 24 hours later, collected the supernatant and measured the inflammatory factors TNF-alpha and IFN-beta, extracted mRNA and measured the expression of TLR3, TLR4 and other correlated indexes of the downstream, analyzed and evaluated Shensu Yin's substance basis of pharmacodynamic actions. RESULT: Shensu Yin drug serum depressed the expression of TLR4, MyD88, TRAF-6, TRAM and TRIF mRNA, as a result, it decreased the amount of TNF-alpha and IFN-beta. CONCLUSION: Depressing the expression of TLR3, MyD88, TRAM and TRIF mRNA may be the elementary basis of Shensu Yin to play heat-clearing and detoxicating effect.