摘要
干扰素α是临床治疗慢性乙型肝炎的一线药物,但临床治疗应答不佳,因此,筛选和鉴定调控干扰素α抗乙肝病毒活性的宿主因子具有重要的意义。本研究首先利用质谱筛选出与干扰素受体1(type I IFN receptor 1,IFNAR1)相互作用的宿主因子聚腺苷二磷酸核糖聚合酶1[Poly(ADP-ribose)polymerase-1,PARP1],采用Co-IP验证。进一步通过荧光定量PCR(qPCR)、双荧光素酶实验和Western blot检测宿主因子PARP1对IFNAR1表达以及干扰素信号通路关键分子表达的调控作用,最后通过基因沉默和过表达策略研究了PARP1对HBV复制以及干扰素α抑制HBV复制的影响。结果显示:通过质谱和Co-IP验证了PARP1和IFNAR1的相互作用;在Huh7.0细胞中,PARP1上调IFNAR1的蛋白水平,促进STAT1和STAT2的磷酸化,从而上调干扰素信号通路下游的干扰素刺激应答元件(IFN-stimulated response elements,ISRE)启动子转录活性以及干扰素诱导的抗病毒基因ISG15,ISG56,APOBEC3G和SAMHD1的表达,促进了干扰素α的抗乙肝效应。本实验表明宿主因子PARP1可以上调IFNAR1表达,促进STAT1和STAT2的磷酸化,进一步加强干扰素α抑制乙肝病毒的效应。
Interferonα(IFN-α)is a first-line drug approved for the treatment of chronic hepatitis B(CHB)infection.However,IFN-αtherapy leads to a limited response against CHB,so investigating the host factors that affect the antiviral activity of IFN-αis important.We identified,using mass spectrometry and co-immunoprecipitation methods,that the host factor Poly(ADP-ribose)polymerase 1(PARP1)interacted with type I interferon receptor 1(IFNAR1).Furthermore,the effects of PARP1 on IFNAR1 expression and the IFN-αsignal-transduction pathway were detected by quantitative polymerase chain reaction(qPCR),dual luciferase assay and Western Blotting in Huh7.0 cells.Finally,the effects of silencing PARP1 expression by small interfering(si)RNA and overexpression of PARP1 on HBV replication and the IFN-αmediated anti-HBV response were detected by qPCR and enzyme-linked immunosorbent assay.These results showed that PARP1could interact with IFNAR1.PARP1 facilitated the expression of IFNAR1,ISG15,ISG56 and APOBEC3G,and phosphorylation of signal transducers and activators of transcription 1(STAT1)and STAT2.Furthermore,PARP1 promoted the antiviral function of IFN-αsignificantly.We showed that the host factor PARP1 could upregulate IFNAR1 expression,promote phosphorylation of STAT1 and STAT2,and further enhance the inhibitory effect of IFN-αon the hepatitis B virus.
作者
熊静
王卓
胡源
XIONG Jing;WANG Zhuo;HU Yuan(Key Laboratory of Molecular Biology on Infectious Disease,Ministry of Education,Chongqing Medical University,Chongqing 400016,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2022年第5期1117-1126,共10页
Chinese Journal of Virology
基金
国家重点研发计划重点专项(项目号:2018YFE0107500),题目:输血传播病毒性肝炎转化医学及精准治疗策略研究
重庆市渝中区科技项目(项目号:20200122),题目:多胺代谢抑制剂DFMO抑制乙肝病毒复制的机制研究。
