Enzyme Hydrolysis Velvet Antlers Peptides And Proliferative Activity

After the native effective components of fresh velvet antler were extracted with ethanol,the resulting remainder slag was hydrolyzed by alcalase, an industrial alkaline protease, togive the total peptides(VATP). The protein content of the hydrolysis product of theremainder slag was5times of that of ethanol-extracted solution from fresh velvet antler.VATP was then separated with Sephadex G-25column to obtain two peaks named VAP-Iand VAP-II. Finally, the main peak VAP-II was further purified by preparative HPLC,resulting two active peptide components (VAP-II-A and VAP-II-B). The molecular weightdistribution of the high active peptide component VAP-II-A was in a rage of500-700Da.The experimental results of MTT and flow cytometry indicate that a high active peptidecomponent, VAP-II-A could stimulate proliferation of a human osteosarcoma cells.VAP-II-A is also able to enhance the average content of DNA in S-phase. And also the alkaliphosphatase (ALP) activity in osteosarcoma cells enhanced obviously with the increase ofVAP-II-A dosage and treatment time. The results are consistent with cell cycle variation andALP level as a marker of bond formation during osteocyte proliferation and differentiation,as well as maturity. Further more, it was found that compared with the control group, therewas not considerable change in cell activity and cell cycle for the VAP-II-A group after thecell pathway was blocked by p38MAPK inhibitor. In addition, we observed that the relativeprotein contents of expressed by BMP, P21and（Cyclin）D1mRNA increased by7times forthe VAP-II-A group compared with the control group, resulted from the SYBR Green Ⅰchimeric fluorescence and RT-PCR experiments.The results obtained in this study provide a clue to explore a new peptide medicinefrom velvet antlers for therapy of osteoporosi.