Screening And Mechanism Of Active Peptides Promoting Cell Migration Based On Hsp90 And LRP-1 Structures

Wound healing is a physiological repair process involving growth factors,cytokines and related tissue cells.Abnormal wound healing,such as excessive healing to form scars or chronic wounds to form ulcers,can cause great harm to the body.Cell migration is the rate-limiting event in wound healing and plays an important role in the wound healing process.When tissue is damaged,cells can release Hsp90? to promote cell migration,transmit cell motility signals through the transmembrane protein LRP-1,and accelerate wound repair.Stomach ulcer is a common digestive system disease,and its repair is a complex wound healing process.At present,the treatment of peptic ulcer mainly focuses on proton pump inhibitors(PPIs)drugs,but there are cases of overuse,serious side effects,and high recurrence rate after drug withdrawal.Based on this situation,it is very urgent to develop a new type of wound-healing drug with good efficacy and low toxicity.Food-derived small molecule polypeptides come from a wide range of sources and have unique advantages,such as being more easily absorbed by the body and having almost no toxic side effects.Not only do they have nutritional value,they are also biologically active.Egg white polypeptide is a food-derived polypeptide prepared from egg white powder.Its various functions have been studied,but there are few reports on cell migration and wound repair.Therefore,it is of great significance to explore egg white polypeptides that can accelerate wound repair.Preliminary work in the laboratory has demonstrated that egg white peptides KR-9(KPHAEVVLR),LN-6(LRDILN),and IL-6(INFEKL)can promote skin wound healing.On this basis,we further study the effect of egg white peptides on other tissues or organs of wound repair.First,the SRL-PFAC technology was used to target the Hsp90/LRP-1 complex,and at the same time,the cell migration assay was used to simultaneously screen the peptide sequences that promote cell migration,and the peptides SE-6(SRLYAE),SS-6 and SS-8(SAESLKIS),LR-6(LYAEER)and RE-7(RYPILPE),further use gradient concentrations of peptides to do cell migration and scratch wound healing experiments to verify the effect of peptides on promoting migration.Next,we performed MTS cell viability experiments with corresponding cell lines to verify the safety of the drug and also to confirm whether the drug would cause excessive cell proliferation and excessive wound healing.Afterwards,we used western blotting experiments to study the mechanism of action of peptides in promoting cell migration and proliferation.After the preliminary in vitro experiments were completed,in order to verify whether the peptides can promote the ulcer repair in gastric ulcer mice,we selected the peptide SS-8 with a strong ability to promote migration,and further proved its effect through in vivo experiments.We established a mouse model of gastric ulcer and administered it for treatment.Through the change of mouse body weight,gastric histomorphological observation,HE staining pathological analysis and determination of serum IL-6 level,the differences between mice in each group were compared,and the healing of gastric ulcers in mice was analyzed from different dimensions,and the effect of SS-8 on ulcer healing was evaluated.Based on the above research,this paper screened out four egg white polypeptides that can promote cell migration,namely SS-8,SE-6,LR-6 and RE-7,which provide scientific theoretical support for the future research and development of drugs for wound healing.At the same time,it also opens up a new road for the development and utilization of egg white powder polypeptides as functional health products in the future.