Study On The Design,Synthesis And Activity Of Velvet Antler Polypeptides

Background and Objective:Velvet antler is one of the famous Traditional Chinese Medicine?TCM?.In the past few years,peptides and protein ingredients were extracted and isolated from velvet antlers,which exerts cell proliferation stimuli in bone,epithelium,blood vessels and neural cells.However,it is difficult to isolate peptides and protein and analyze their effective ingredients,for example,complex separation and purification process,long extraction time,easy deactivation,more consumable raw materials and low recovery rate.Therefore,the research on medicinal value and marketing of velvet antler were restricted.In this work,a 14-amino acid peptide and its four peptide fragments were synthesized based on peptides isolated from velvet antlers that previously reported.And then,the protective effect against 6-hydroxydopamine?6-OHDA?induced cell death were evaluated in human neuroblastoma cell line SH-SY5Y to develop the potential therapeutic agent for Parkinson's disease?PD?treatment.Methods:?1?Five peptides,including 14-amino acid peptide EP₁₄ and its four peptide fragments EL₅,DL_5,EL₁₀ and DP₉,were synthesized using mixed anhydride method.And then,they were analyzed and identified by high performance liquid chromate-graphy?HPLC?and electrospray ionization mass spectrometry?ESI-MS?.?2?The SH-SY5Y cells were incubated with 300?mol/L 6-OHDA to establish PD model.The protective groups were treated with different concentration of five peptides respectively.Viability of SH-SY5Y cells were measured using CCK-8 assay.Cell apoptosis was detected using Hoechst 33258 staining and Annexin V-FITC/PI flow cytometry.Results:?1?Five target peptides were designed rationally and synthesized successfully.?2?Compared with the model group,viability of SH-SY5Y cells enhanced while late apoptotic rate reduced significantly?P<0.05?in target peptides groups DL5,EL10 and EP₁₄4 according to CCK-8 and Annexin V-FITC/PI flow cytometry results.?3?Hoechst33258 staining showed that nuclear condensation and fragmentation of target peptides groups DL₅,EL₁₀ and EP₁₄ were significantly alleviated.Conclusion:Synthesized peptides DL₅,EL₁₀0 and EP₁₄,originated from velvet antlers can protect against 6-OHDA-induced neural cell death by inhibiting cell apoptosis.