不同类型干扰素对Toll样受体在Huh7细胞中表达的影响

The impact of different types of interferon on the expression of Toll-like receptors in Huh7 cells

目的探讨不同类型干扰素对肝细胞Toll样受体(toll like receptor,TLR)3、4、7、9表达的影响。方法感染丙型肝炎病毒(hepatitis C virus,HCV)JFH-1病毒的Huh7细胞分别加入干扰素α(interferon,IFN-α)、IFN-β、IFN-γ、IFN-λ1处理24 h,以实时定量聚合酶链反应(real time polymerase chain reaction,RT-PCR)分析Toll样受体3、4、7、9以及HCV复制水平的变化。结果各干扰素均能抑制HCV的复制(t分别为-45.699、-68.165、-70.994、-67.111,均有P<0.001)。方差分析显示各干扰素处理组间TLR3、TLR4、TLR7及TLR9的相对表达量差异有统计学意义(F分别为125.901、263.537、163.289、31.774,均有P<0.001),进一步两两比较可知,IFN-α、IFN-β能上调TLR3(t分别为21.312、25.420,均有P<0.001)、TLR7(t分别为26.456、24.289,均有P<0.001)的表达,同时下调TLR9的表达,而对TLR4的表达没有影响;IFN-γ可上调TLR4、TLR7的表达,对TLR3和TLR9的表达没有影响;IFN-λ1可上调TLR3、TLR7、TLR9的表达水平,但对TLR4的表达没有影响。结论不同干扰素对肝细胞不同Toll样受体表达的作用不同,反映出不同干扰素具有不同的抗病毒作用机制。

Objective To investigate the effects of different types of interferon on the expression of TLR3 ,TLR4,TLR7 and TLR9 in. human hepatocytes. Methods HCV JFH-l-infected Huh7 cells were treated by IFN-α, IFN-β, IFN-γ, IFN-λ1 respectively for 24 h. The expression of TLR3, TLR4, TLR7 and TLR9 of Huh7 cells and the levels of HCV RNA were determined by real time PCR. Results All interferons （IFN-α, IFN-β, IFN-γ, IFN-λ1 ） could significantly inhibit HCV replication in Huh7 ceils （t = - 45. 699, - 68. 165, - 70. 994, - 67.111 respectively, all P 〈 0. 001 ）. The ANOVA showed that the levels of TLR3, TLR4, TLR7 and TLR9 expression among different IFN - treated groups had significant difference （ F = 125.901,263. 537,163. 289,31. 774 respectively, all P 〈0. 001 ）, and comparisons between two groups showed that IIFN-α and IFN-β treatments significantly increased the expression of TLR3, TLR7 （t = 21. 312, 25.420, 26. 456, 24. 289, all P 〈 0. 001 ）, and decreased the expression of TLR9, but had no significant effect on the expression of TLR4. IFN-γ treatment significantly increased the expression of TLR4 and TLR7, but had no significant effect on the expression of TLR3 and TLR9. IFN-λ1 treatment increased the expression of TLR3, TLR7 and TLR9, but had no significant influence on the expression of TLR4. Conclusions Different types of interferon have different effects on TLR expression pattern in human hepatocytes, implying that antiviral mechanisms vary with the types of interferons used against HCV infection in hepatocytes.