Research On The Extraction Isolation And Expression Of Active Components From Pilose Antler

In order to elucidate the biochemical mechanism of therapeutic effects of pilose antler, the extraction of freeze dried pilose antler powder was fractionated by a combination of chromatography of different chemistries such as gel filtration chromatography on Sephacryl S-200HR, ion-exchange chromatography on SOURCE 15Q, reverse phase chromatography on C18 and chromatography on Superdex peptide 10/300GL. As a result two bioactive components, P2-3 and b were obtained.P2-3, electrophoretically pure, with a molecular of 53.3 kDa, were demonstrated to be capable of increasing the proliferation rate of both Madin-Darby canine kidney cells (MDCK) and mice spleen cells, with a similar function pattern. With the increase of P2-3 concentration, the proliferation rate of MDCK cells increased at first, and then decreased when the concentration of P2-3 exceeded a certain value, with the maximum of 73.0% and 63.3% increase for MDCK cells and spleen cells, respectively at the concentration of 0.124mg P2-3/ml. The isoelectric point of P2-3 fraction is 5.4, as was measured by ion-exchange chromatography.b fraction, which was proved to be pure by capillary reverse phase chromatography, was demonstrated to also promote the proliferation of both MDCK cells and mice spleen cells. MDCK cell proliferation was inhibited at low concentration. With the increase in its concentration, MDCK cell proliferation was enhanced to reach the maximum of 64.3% of the control group at a concentration of 11.1 μg/ml. The effect of b fraction on mice spleen cell proliferation was less potent than on MDCK cells, with the maximum of 34.9% at 11.1μg/ml concentration.Since cell proliferation and differentiation constitute a major process in wound healing, the isolation P2-3 and b from pilose antler and the identification of their bioactivity of increasing the proliferation of epithelial cells, have provided another molecular clue to the elucidation of pilose antler's therapeutic effect in wound healing, as well as to the development of a new wound healing drugs from pilose antler. Consisting of various immune cells, spleen plays an important role in the immune...