Optimization Of Extraction Process Of Velvet Antler Polypeptides And Study On The Effects Of Anti-oxidation And Promoting Healing

Velvet antler is the unossified and fuzz young horn of sika deer or red deer.It contains phospholipids,sugar lipids,fatty acids,proteins and other components,and has many pharmacological activities such as immune regulation,cancer prevention,anti-osteoporosis and anti-inflammatory.In recent years,due to the wide application of natural peptides in skin anti-aging products,more and more attention has been paid to natural peptides.Therefore,this study was based on the extraction of bioactive peptides from velvet antler.The antler polypeptides which can promote the proliferation and collagen secretion of mouse fibroblasts(NIH/3T3)were obtained by using the antioxidant activities,cell proliferation and collagen secretion of different molecular weight peptides as screening conditions.And the mechanism study was carried out.The aim was to improve the extraction rate of velvet antler polypeptides,reflect the medicinal value of velvet antler,and have guiding significance for the further development and utilization of velvet antler.The main research contents and results were as follows:1、The polypeptides in velvet antler were extracted by ultrasonic extraction method,and the optimal extraction process was obtained by single factor and response surface optimization experiments: the ultrasonic duration was 46 min,the pH was 5.8,the solid-liquid ratio was 15 times,and the ethanol concentration was 46%.2、The 1-3kDa,3-5kDa and 5-10 kDa velvet antler peptides were obtained by membrane separation with antler polypeptides desalted with sephadex G-25,and their molecular weight was determined by MALDI-TOF-MS.3、The activity of velvet antler polypeptides was detected by antioxidant,NIH/3T3 proliferation and collagen secretion assay,respectively.The results showed that,compared with the blank group,1-3kDa velvet antler polypeptide had the best antioxidant capacity,3-5kDa velvet antler polypeptide had the best effect in promoting NIH/3T3 cell proliferation,and 5-10 kDa velvet antler polypeptide had the best effect in promoting collagen secretion.4、Effect of NIH/3T3 on proliferation and collagen secretion of velvet antler polypeptide 1-10 kDa and its mechanism were studied.Compared with the control group,1-10 kDa velvet antler polypeptide can promote the proliferation of NIH/3T3 cells,promote the secretion of collagen and the healing of cell scratches.The expressions of p-ERK1/2 and TGF-β1 in NIH/3T3 cells were increased by Western blotting and immunofluorescence,suggesting that the mechanism may be achieved by activating the phosphorylation of p-ERK1/2 and the expression of TGF-β1.