Rao Hui-Ying, Wang Jiang-Hua, Liu Feng, Fei Ran, Liu Zhi-da, Wei Lai
Peking University People's Hospital, Peking University Hepatology Institute, Beijing, China.
Zhonghua Gan Zang Bing Za Zhi. 2007 Dec;15(12):897-901.
To further study the mechanism of the inhibitory effect of interferon beta-1a (IFN beta-1a) on the activation of human hepatic stellate cell (HSC) LX-2, and to analyze the differences on the protein expression in LX-2 induced by I IFN beta-1a.
Cultured LX-2 cells were treated with 2000 U/ml IFN beta-1a for 48 h. Two-dimensional gel electrophoresis (2-DE) was performed to compare protein patterns of the control (untreated) and IFN beta-1a treated LX-2 and for quantitative and qualitative analyses of protein expression. A rat liver fibrosis model was established and the rats were sacrificed and their various tissues were obtained for the same analyses. Western blotting and RT-PCR were used to validate the expression of the changed proteins after treatment of IFN beta-1a in LX-2 cells and of various tissues of the rats.
708 +/- 25 spots were detected in control LX-2 cells and 804 +/- 32 spots in IFN beta-1a-treated LX-2 cells. A match rate of 73%-82% was achieved. The results also showed that 31 protein spots displayed quantitative changes in expression after IFN beta-1a treatment. Of the 31 spots, 21 proteins were identified, of which, one was newly found, two were enhanced in abundance and 18 showed lower expressions. The newly found protein was glia maturation factor beta (GMF beta). The treatment of LX-2 with IFN beta-1a increased the production of GMF beta(GMF beta) protein in comparison with the untreated cells (t=1.81, P < 0.01). The expression of GMF beta protein (1.81 vs 0.10) and mRNA (0.85 vs 0.12) were more in the normal liver tissues than in the cirrhotic liver tissues (t=2.53, 2.13 respectively, P < 0.01). The expressions of GMF beta protein and mRNA were weak in rat heart and lung tissues, however, they were strong in rat liver, kidney, spleen and brain tissues (t=1.91, 1.94 respectively, P < 0.01).
There is a significant difference of protein expression levels between IFN beta-1a untreated and treated LX-2 cells. These proteins, especially GMF beta, may be involved in an inhibition process of IFN beta-1a on activation and apoptosis of LX-2 cells. This proteome study may be useful in further studies of the relationship of IFN beta-1a treatment and human liver diseases.
进一步研究β-1a干扰素（IFNβ-1a）抑制人肝星状细胞（HSC）LX-2活化的机制，并分析IFNβ-1a诱导LX-2细胞中蛋白质表达的差异。
用2000 U/ml IFNβ-1a处理培养的LX-2细胞48小时。进行二维凝胶电泳（2-DE）以比较对照（未处理）和IFNβ-1a处理的LX-2细胞的蛋白质图谱，并对蛋白质表达进行定量和定性分析。建立大鼠肝纤维化模型，处死大鼠并获取其各种组织进行相同分析。采用蛋白质印迹法和逆转录-聚合酶链反应（RT-PCR）验证IFNβ-1a处理后LX-2细胞及大鼠各种组织中变化蛋白质的表达。
对照LX-2细胞中检测到708±25个斑点，IFNβ-1a处理的LX-2细胞中检测到804±32个斑点。匹配率为73%-82%。结果还显示，31个蛋白质斑点在IFNβ-1a处理后表达出现定量变化。在这31个斑点中，鉴定出21种蛋白质，其中一种是新发现的，两种丰度增加，18种表达降低。新发现的蛋白质是胶质细胞成熟因子β（GMFβ）。与未处理的细胞相比，用IFNβ-1a处理LX-2可增加GMFβ蛋白的产生（t=1.81，P<0.01）。正常肝组织中GMFβ蛋白（1.81对0.10）和mRNA（0.85对0.12）的表达高于肝硬化肝组织（分别为t=2.53,2.13，P<0.01）。GMFβ蛋白和mRNA在大鼠心脏和肺组织中表达较弱，然而，它们在大鼠肝脏、肾脏、脾脏和脑组织中表达较强（分别为t=1.91,1.94，P<0.01）。
未处理和经IFNβ-1a处理的LX-2细胞之间蛋白质表达水平存在显著差异。这些蛋白质，尤其是GMFβ，可能参与IFNβ-1a对LX-2细胞活化和凋亡的抑制过程。这项蛋白质组学研究可能有助于进一步研究IFNβ-1a治疗与人类肝脏疾病的关系。
