绿原酸通过调节TLR3-TRIF通路抑制小胶质细胞炎性反应的机制研究

Mechanism of Chlorogenic Acid Inhibiting Inflammatory Response of Microglia by Regulating TLR3-TRIF Pathway

目的:研究绿原酸(chlorogenic acid,CGA)在单纯疱疹病毒1型(herpes simplex virus-1,HSV-1)感染的BV2小胶质细胞中的抗炎作用。方法:把不同浓度梯度的CGA加入到正常BV2细胞中,通过CCK-8试剂盒测定CGA最大无毒浓度范围,以HSV-1感染BV2细胞建立HSV-1细胞模型,然后用不同浓度梯度的CGA进行处理,通过Real-time PCR检测Toll样受体(toll-like receptor 3,TLR3)信号通路中β干扰素TIR结构域衔接蛋白(TIRdomain-containing adaptor inducing interferon-β,TRIF)mRNA的表达,Western blot检测TLR3、TRIF的蛋白表达情况,ELISA检测炎性因子组织细胞内干扰素α(interferon-α,IFN-α)水平。结果:单纯疱疹病毒性脑炎细胞模型中TLR3及其下游分子TRIF的mRNA、蛋白表达及IFN-α水平均明显高于正常对照组(P<0.05);而绿原酸干预可有效抑制炎性反应,TLR3及其下游分子TRIF的mRNA、蛋白表达及IFN-α水平均明显降低。结论:绿原酸能明显抑制小胶质细胞内TLR3通路,降低HSV-1引起的炎性反应。

Objective To study the anti-inflammatory effects of chlorogenic acid(CGA)in BV2 microglia infected with herpes simplex virus-1(HSV-1).Methods Chlorogenic acid with different concentration gradient was added to normal BV2 cells.The maximum non-toxic concentration range of CGA was determined by CCK-8 kit.The HSV-1 cell model was established using BV2 cells infected with HSV-1,and then treated with CGA with different concentration gradients.The mRNA expression of TLR3 and its downstream signal molecule TRIF was detected by Real-time PCR,and the protein expression of TLR3 and TRIF was detected by western blot.ELISA was used to detect the level of IFN-αin inflammatory tissue.Results The mRNA and protein expression of TLR3 and its downstream molecule TRIF and the level of IFN-αin HSV encephalitis cell model were significantly higher than those in normal control group(P<0.05).Chlorogenic acid intervention could effectively inhibit inflammatory reaction.The mRNA and protein expression of TLR3 and TRIF and the level of IFN-αwere significantly decreased.Conclusion Chlorogenic acid can significantly inhibit the TLR3 pathway in microglia and reduce the inflammatory reaction induced by HSV-1.