Research On Biologically Active Peptide Stimulate Proliferation And Differentiation Of Cells And Other Related Funtions

Ejiao (Colla Corii Asini) as an important traditional Chinese medicine has a lot offunctions. We found donkey serum albumin is a major protein component in Ejiao, and inthis research, we mainly study the biologically active peptide HP-6which was originatedfrom donkey serum albumin. Through testified the activity of HP-6, a new method ofpharmacological research on Ejiao was put forward and theory basis of its applicationwas built. Based on the experiment results of HP-6, the further study was focus ondonkey serum which is rich in donkey serum albumin. Preliminary research onapplication of donkey serum was performed by investigated the effects of donkey serumhydrolysates.The influences of HP-6on mouse bone marrow cells, HL-60and HepG-2wereinvestigated in vitro. The results showed as follows:1. HP-6could stimulate theproliferation of mouse bone marrow stromal cells (mBMSCs), and the best concentrationwas0.15μmol·L-1with reproduction rate at (63.52±3.94)%. The influence on mousebone marrow suspension cells was tested, and the best concentration was0.015μmol·L-1with reproduction rate at (24.66±3.70)%;2. Observed by confocal laser scanningmicroscope, HP-6could go through the cell membrane and get into cytoplasm;3. HP-6could not stimulate but somehow inhibit the proliferation of HL-60, HepG-2and HepG-2colony;4. Low concentration of HP-6got a better influence on stimulate mouse bonemarrow CFU-E colony.Hemorrhage anemia mouse model and anemia mouse model induced bycyclophosphamide were established to investigate the effect of HP-6in vivo. The resultseems that HP-6could enhance mouse hemopoiesis function and the resistance tochemotherapeutic injury. In vivo research also indicated that HP-6could enhance thedevelopment of liver and lung in mice and without long-term biotoxicity.The donkey serum was hydrolyzed by trypsin under different conditions, and thebest conditions were pretreatment temperature at85℃, initial pH at9, hydrolysistemperature at55℃, enzyme amount at2500U·mL-1, and hydrolyzed for9h. However, cell proliferation assays in vitro indicated the hydrolysates obtained at5and7h hadbetter function on mBMSCs proliferation than hydrolysates obtained from other times. Invivo analysis indicated that donkey serum protein was safe to mice and could influencethe mice growth especially the liver growth. The hydrolysates prepared by reacting withtrypsin in vitro had a little less effect on mice growth compared with hydrolysates reactedwith inactivated trypsin. However, the hydrolysates prepared by reacting with trypsin invitro had more effect on the development of lung.Above all, bioactive peptide HP-6obtained from donkey serum albumin bybioinformatics’ way had the same functions as Ejiao in hematopoietic system, and donkeyserum which is rich in donkey serum albumin also had similar functions as HP-6afterhydrolyzed.