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

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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.