IGFBP-rP1对肝星状细胞活化及核因子-κB活性的影响  

Effect of IGFBP-rP1 on hepatic stellate cells and the activity of nuclear factor-κB

作　　者：刘毅[1] 刘立新[1] 郭晓红[1] 张骞骞[1] 

机构地区：[1]山西医科大学第一临床医学院

出　　处：《中国病理生理杂志》2008年第9期1789-1793,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.30740031);教育部新世纪优秀人才支持计划资助项目(No.NCET-06-0264)

摘　　要：目的:明确胰岛素样生长因子结合蛋白相关蛋白1(ⅠGFBP-rP1)是否具有活化肝星状细胞(HSC)、增加细胞外基质(ECM)合成的作用;并探讨可能的信号转导途径。方法:大鼠肝星状细胞株(HSC-T6)体外培养,分别设立空白对照组(加入等量PBS)和不同浓度的ⅠGFBP-rP1处理组,干预因素处理24 h后收集细胞爬片或制备单细胞悬液,采用免疫细胞化学染色观察HSC-T6中α-平滑肌肌动蛋白(α-SMΑ)、Ⅰ型胶原(colla-gen Ⅰ)、纤维连接蛋白(FN)的表达变化;流式细胞仪检测HSC-T6中核因子-κB p65(NF-κB p65)的DNΑ结合活性。结果:免疫细胞化学染色结果发现,ⅠGFBP-rP1各处理组α-SMΑ、collagen Ⅰ、FN的表达均较空白对照组显著增强,且在一定范围内呈剂量依赖关系;流式细胞仪检测结果显示,ⅠGFBP-rP1各处理组NF-κB p65阳性细胞百分数均较空白对照组明显增高。结论:ⅠGFBP-rP1可以激活HSC,且在一定剂量范围内随着ⅠGFBP-rP1剂量的增加HSC活化的程度逐渐增强;ⅠGFBP-rP1可使ECM的重要组成成分collagen Ⅰ和FN的合成增加;ⅠGFBP-rP1可增强HSC中NF-κB p65的DNΑ结合活性。AIM ： To identify the effect of insulin - like growth factor binding protein - related protein 1 IG- FBP - rP1 ） on the activation of hepatic stellate cells ,（HSC） and the synthesis of extracellular matrix （ ECM ）, and to investigate the possible pathway of signal transduction. METHODS ： Rat HSC - T6 was cultured in vitro and established respectively the control group （ incubated with PBS） and the groups treated with different concentrations of IGFBP - rP1. Cell - coated dishes were attained, and the single cell suspension was prepared after 24 h, then the expression of alpha - smooth muscle actin （α - SMA）, collagen I and fibronectin （FN） were detected by immunocytochemical staining. The DNA binding activity of NF - κB p65 was detected by flow cytometry. RESULTS ： The results of immunocytochemical stai- ning showed that the positive stainings of α - SMA, collagen I and FN in the groups of IGFBP - rP1 were significantly higher than those in control group, and a dose - dependent relationship in a certain range was observed. Flow cytometry analysis showed that the percentage of NF - κB p65 positive cells in the groups of IGFBP - rP1 markedly increased compared with control group. CONCLUSION ： HSC is activated by IGFBP - rP1 and the level of HSC activation gradually enhances in a certain dose range of IGFBP - rP1. The synthesis of both collagen I and FN is increased by IGFBP - rP1. IGFBP - rP1 en- hances the DNA binding activity of NF - κB p65 in HSC.

关 键 词：胰岛素样生长因子结合蛋白质相关蛋白质1 肝星状细胞 细胞外基质 NF—κB 

分 类 号：R363[医药卫生—病理学]