Preparation Of Polypeptides And Collagen Peptides From Sika Deer And Preliminary Study On Inhibiting Proliferation Of Breast Cancer Cells

The antler plate is the ossification material of red deer and antler remaining in the staghorn after red deer or spotted deer are sawed off by the deer antler.It falls off on its own in the spring before the new antler grows.It is shaped like a plate,so it is called an "antler plate",which is the main raw material for antler glue and antler cream.Antler plate is a by-product of velvet antler.For a long time,people neglected the biological activity of antler plate,resulting in the protein resources of antler plate failing to reflect its medicinal value,resulting in the economic value of antler plate failing to be fully reflected,The protein and polypeptide play an important role in life activities.The main pharmacological active ingredient of antler plate is approximately 40% of crude protein.However,a large number of studies have confirmed that proteins are absorbed in the form of small molecule polypeptides in vivo.Therefore,this study aims to extract biologically active polypeptidesand collagen peptides from the antler dish and conduct preliminary studies on the inhibition of the proliferation activity of human breast cancer cell MCF-7 in order to improve the utilization of the antler disk and to highlight the medicinal value of the antler plate.It is of guiding significance to the further research and development and utilization of the precious antler dish.The main research contents and results are as follows:In this experiment,the secondary ground antler dish powder was used as the raw material,and combined with weak acidic buffer salt solution and ultrasonic assisted technology to extract.Sephadex G-25 molecular sieve chromatography was used for salt removal,and Sephadex G-50 molecular sieve was used to obtain the sika deer dish.Molecular protein;On the other hand,the Sika deer horn collagen hydrolyzed peptide was obtained for 4 extractions with 100°C hot water bath,papain and trypsin complex enzymolysis.,.The small molecule proteins and collagen hydrolyzed peptides wasanalyzed by using quinoline dicarboxylate method,Tricine-SDS-PAGE electrophoresis and mass spectrometry.On this basis,the human breast cancer cell line MCF-7 was used as the in vitro research object.The inhibition of the proliferation of human breast cancer cell line MCF-7 by total peptide,collagen,small molecule protein and collagen hydrolyzed peptide was detected by MTT assay.The results showed that the higher the ultrasonic power,the higher the total peptide concentration of sika deer horn disk protein.Combined with the result of SDS-PAGE electrophoresis,we selected the total peptide crude material after ultrasonic treatment for 200 watts to continue the follow-up test;Sika deer horn disk small molecule protein via Tricine-SDS PAGE three-dimensional gel electrophoresis detection of its molecular mass concentrated in the following 6500Da;composite enzymatic hydrolysis of collagen peptides by electrophoresis and mass spectrometry detection of their molecular mass concentrated in 6500 Da below;The inhibition of the four cell proliferation found that all the proliferation of human breast cancer cell line MCF-7 was significantly inhibited.The total peptide and collagen concentration of 30 ?g/mL had the highest inhibition rate after 72 hours;the concentration of small molecule protein and collagen hydrolyzed peptide at 20 ?g/mL after 72 hours had the highest inhibition rate.The inhibition of MCF-7 cell proliferation was compared: small molecule protein> collagen hydrolyzed peptide> total peptide> collagen.