Screening for biologically active compounds which influence the formation of collagen fibers using culltured liver M cells

从大鼠肝脏建立的细胞系具有独特的特征，尽管它是细胞系，但它产生胶原蛋白样分化细胞。胶原蛋白的过度形成可能导致肝纤维化，并且当从伤口恢复时，预期胶原蛋白的快速形成。在这个项目中，首席研究员打算从天然来源中分离出影响培养的肝细胞中胶原蛋白形成的生物活性化合物。经过对许多植物的粗甲醇提取物的广泛筛选，几种提取物显示出相当大的减少胶原纤维形成的活性。这种植物之一是豆腐柴，收集在冲绳县。将活性转移到正丁醇可溶性部分，然后通过高孔合成树脂（HP-20）、硅胶、液滴逆流和高效液相色谱对甲醇乙酯的正丁醇可溶性部分进行分析指导分级，最后分离出纯的活性化合物，NMR光谱分析表明其为苯乙醇的衍生物，例如广泛从植物来源中分离的类叶升麻苷。

A established cell line from rat liver has a unique feature that although it is the cell line, it produces collagen protein like differentiated cells. Excessive formation of collagen protein may cause liver fibrosis and when recovery from wound, rapid formation of collagen protein is expected. In this project, the head investigator intended to isolate biollogically acitive compounds which influence for formation of collagen protein in cultured liver cells from natural sources.After extensive screening of crude MeOH extracts of many plants, several extracts showed considerable activities to reduce the formation of collagen fibers. One of such plants was Premna subscandens, collected in Okinawa Prefecture. The activity was transferred to n-BuOH soluble fraction and then active principles were pursued by assay guided fractionation of the n-BuOH solable fraction of the MeOH estract by highly porous synthetic resin (HP-20), silica gel, droplet counter-current and high-performance liquid chromatographies.Finally active compound was isolated as a pure state and NMR spectroscopic analyzes revealed that it is a derivative of phenylethanoid, such as acteoside which has been isolated widely from plant sources.