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目的研究干扰素λ1(interferonλ1,IFN-λ1)对肝细胞内自噬相关基因表达的影响,探讨IFN-λ1抗HCV的新机制。方法感染HCV JFH-1毒株的Huh7细胞加入IFN-λ1处理,采用实时荧光逆转录定量PCR法分析细胞自噬相关基因LC3、Beclin、ATG3、ATG5、ATG7、Bcl-xl、Bad mRNA表达以及HCV RNA水平的变化,采用Western Blot检测自噬相关基因和HCV Core在蛋白水平的变化。结果 IFN-λ1抑制HCV在Huh7细胞中的复制,与对照组相比,IFN-λ1可以下调HCV RNA的水平(t=-4.513,P=0.011)和HCV Core蛋白相对表达量(t=6.943,P=0.020);IFN-λ1可下调细胞自噬基因ATG3、ATG5的mRNA表达(t=10.311,P=0.007;t=-14.246,P=0.004),同时上调细胞自噬抑制因子Bcl-xl的mRNA表达(t=-5.246,P=0.034)。IFN-λ1对ATG3、ATG5的蛋白表达同样具有抑制作用(t=6.668,P=0.022;t=13.343,P=0.006)。结论干扰素λ1可能通过抑制肝细胞内自噬基因ATG3、ATG5的表达以及上调细胞自噬抑制因子Bcl-xl的表达,破坏细胞自噬过程,从而达到抗HCV的作用。
Objective To study the effect of interferon λ1 (IFN-λ1) on the expression of autophagy-related genes in hepatocytes and to explore the new mechanism of IFN-λ1 anti-HCV. Methods Huh7 cells infected with HCV JFH-1 strain were treated with IFN-λ1 and the expressions of LC3, Beclin, ATG3, ATG5, ATG7, Bcl-xl and Bad mRNA were detected by real-time fluorescence reverse transcription polymerase chain reaction RNA level changes, Western Blot detection of autophagy-related genes and HCV Core protein level changes. Results IFN-λ1 inhibited the replication of HCV in Huh7 cells. Compared with the control group, IFN-λ1 down-regulated HCV RNA level (t = -4.513, P = 0.011) and relative expression of HCV Core protein P = 0.020); IFN-λ1 down-regulated the mRNA expression of autophagy genes ATG3 and ATG5 (t = 10.311, P = 0.007; t = -14.246, P = 0.004) and up-regulated the expression of autophagy inhibitor Bcl-xl mRNA expression (t = -5.246, P = 0.034). IFN-λ1 also inhibited ATG3 and ATG5 protein expression (t = 6.668, P = 0.022; t = 13.343, P = 0.006). Conclusion Interferon λ1 can inhibit the autophagy of hepatocytes by inhibiting the expression of autophagy genes ATG3 and ATG5 and up-regulating the expression of autophagy inhibitor Bcl-xl.