摘要
干扰素α(IFN-α)是临床最常用的抗乙型肝炎病毒(HBV)药物之一。泛素特异性蛋白酶18(USP18)被证实是抑制IFN-α抗HBV活性的因子,但USP18是否对干扰素λ(IFN-λ)抗HBV有影响还尚未可知。为了明确USP18对IFNλ抗HBV活性的影响,本研究以Hep G2. 2. 15细胞作为乙肝体外模型,采用脂质体转染法分别向细胞转染p EGFP-USP18、PEGFP-N1经48 h,再经IFN-α和IFN-λ处理24 h,分为阴性对照组﹑USP18过表达+IFN-α组﹑空载组+IFN-α组﹑USP18过表达+IFN-λ组﹑空载组+IFN-λ组。采用Western印迹、RT-q PCR和ELISA检测各组的乙肝病毒标志物、STAT1/p STAT1和下游的干扰素刺激基因(ISGs)的表达。结果显示,与阴性对照组和空载组相比,USP18蛋白在过表达组明显升高(P <0. 05),过表达细胞模型构建成功;在IFN-α处理的两组中,空载组中HBs Ag、HBe Ag、HBc Ag及HBV-DNA的表达均低于USP18过表达组,差异有统计学意义(P <0. 05)。而IFN-λ处理组中,乙肝病毒标志物的差异不明显。在IFN-α处理组中,空载组的ISG15、Mx A、IFIT1和p STAT1表达均高于USP18过表达组,差异有统计学意义(P <0. 05),而在IFN-λ处理组中ISGs和p STAT1的表达无明显差异。上述结果证实,USP18可通过抑制JAK/STAT信号通路的激活来减弱IFN-α抗HBV的活性。研究还证实,IFN-λ可发挥抗HBV的作用,USP18不通过JAK/STAT信号通路抑制其抗HBV活性。
Interferon alpha（ IFN-α） is one of the most widely used anti-hepatitis B virus（ HBV）medicine. It has been reported that ubiquitin specific protease 18（ USP18） inhibits the anti-HBV activity of IFN-α,but whether USP18 has an effect on the anti-HBV activity of IFN-λ was unclear. Here we aim to detect the anti-HBV effect of USP18 on IFN-λ in an in vitro model. HepG2. 2. 15 cells were transfected with two different plasmids： the empty vector（ p EGFP-N1） and USP18 overexpressed plasmid（ p EGFPUSP18）,and then were treated with IFN-α and IFN-λ for 24 hours,respectively. Untreated groups served as the negative control. The expression of HBV markers,STAT1/p STAT1 protein expression and interferon stimulated genes（ ISGs） were tested on HepG2. 2. 15 by Western blotting,quantitative realtime PCR（ RT-q PCR） and enzyme-linked immunosorbent assays（ ELISA）. The results showed that USP18 was successfully overexpressed in the overexpression group compared with both negative control and empty vector control groups（ P〈 0. 05）,which demonstrated the successful establishment of vitro models. In IFN-α treated groups,the expression of hepatitis B surface antigen（ HBs Ag）,hepatitis B e antigen（ HBe Ag）,hepatitis B core antigen（ HBc Ag） and HBV-DNA in overexpression groups were significantly higher than empty vectors（ P 〈0. 05）,while there was little difference in the IFN-λtreatment group. In addition,the expression of ISG15,Mx A,IFIT1 and p STAT1 of the empty vector were obviously higher than those in overexpression groups in the treatment group of IFN-α（ P 〈0. 05）,but there was also no significant difference in ISGs and p STAT1 expression in IFN-λ treatment groups.USP18 inhibits the anti-HBV activity of IFN-α by suppressing the activation of the JAK/STAT signaling pathway. In contrast,USP18 has no effect on the anti-HBV activity of IFN-λ via the JAK/STAT signaling pathway.
作者
张小梅
雷青松
秦波
李麟
ZHANG Xiao-Mei;LEI Qing-Song;QIN Bo;LI Lin(Department of Infectious Diseases,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Department of Hepatic Diseases,Chongqing Traditional Chinese Medical Hospital,Chongqing 400021,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2018年第9期954-961,共8页
Chinese Journal of Biochemistry and Molecular Biology
基金
Supported by National Natural Science Foundation of China(No.81271838)~~
关键词
乙型肝炎病毒
泛素特异性蛋白酶18
干扰素Α
干扰素λ
hepatitis B virus （HBV）
ubiquitin specific proteinase 18 （ USP18 ）
interferon-alpha
interferon-lambda （ IFNλ ）
