IGFBP-rP1对肝星状细胞活化及核因子-κB活性的影响

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（collagen 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。

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

AIM:To identify the effect of insulin-like growth factor binding protein-related protein 1(IGFBP-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 staining 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 enhances the DNA binding activity of NF-κB p65 in HSC.