【摘要】 应用亲和层析、凝胶过滤法以及 X型蛋白酶消化结合制备电泳分别提取纯化大鼠 FN及 FN细胞结合片段 (12 0KDa FN - f) ,后者经生物素标记后 ,用亲和细胞化学方法研究大鼠肝星状细胞 (HSC) FN受体 (FNR)表达及调控。结果显示 ,生物素标记的 12 0 KDa FN- f与 FNR的结合具有特异性。原代培养 5 d、 7d的正常大鼠 HSC表达 FNR较培养 1d、 3 d的明显增强。血小板源性生长因子 (PDGF)和转化生长因子 - β1(TGF- β1)可上调大鼠 HSC表达 FNR,全反式维甲酸(at RA)则下调细胞因子再激活的 HSC表达 FNR,并呈剂量依赖性。本文建立了检测 FNR的一种新方法 ,即配体 (12 0 KDaFN- f) -受体 (FNR)亲和细胞化学方法 ,该方法能反映 FNR的总体水平及活性状态。同时初步探讨了影响大鼠 HSC表达FNR的因素 ,确切机制有待进一步研究。更多还原

【Abstract】 In this study, affinity chromatography、 gel filtration and preparative electrophoresis were used to prepare the rat plasma fibronectin and purify the cell binding fragment (120 KDa FN f) from the thermolysin digesting FN solution The purified 120 KDa FN f was labeled with biotin and used to detect the expression and modulation of fibronectin receptor (FNR) in normal rat hepatic stellate cells (HSC) by means of affinity cytochemistry method The results indicated that the specificity of binding Biotin 120 KDa FN f to FNR was believable The expression of FNR in activated HSCs at day 5 and 7 of primary culture was strongly increased in contrast with quiescent HSCs at day 1 and 3 of primary culture Platelet derived growthe factor (PDGF) and transforming growth factor β1 (TGF β1) could up regulate the expression of FNR in ucltured rat HSCs, but all trans retinoic acid (atRA) could down regulate the expression of FNR in cytokine activated rat HSCs in a dose dependent manner This study employed 120 KDa FN f which was prepared by ourselves as a ligand to establish a novel affinity cytochemistry method to detect FNR in HSC The whole level and activity of FNR could be shown using this method The factors affecting the expression of FNR in cultured rat HSCs were also discussed, but the mechanism remains unclear and is worth further study \;更多还原