The Research Of Earthworm Anti Damage Of Effective Components On The Mouse Trauma

Objective:To investigate the effect and mechanism of earthworm anti damage ef fective components(EADEC) in antibiosis, antiphlogosis and the promoting of wound heal ing, we explored the therapeutic effect of EADEC on mechanical injured mice,thus provided a theoretical basis for the therapeutic effect and mechanism research of wou nd healing with EADEC,and also gave us new ideas for the future development of a nimal origin pharmaceutical preparations and extended applications of earthworm extr act in the field of medicine.Methods:1.The EADEC was extracted from earthworm by electric shock,ultrason ic cell disruption,ammonium sulfate salting,ultrafiltration and bacterial sieve.The protei n molecular weight range and antibacterial activity were tested by polyacrylamide gel electrophoresis and susceptibility testing.The EADEC was made into spray with excipi ents such as propylene glycol,Tween 80,sucrose and Nacl.2.60 KM mice that had made dorsal wound were divided into positive control g roup, drug control group and EADEC group and accepted the spraying treatment.Clini cal and local healing situation were observed everyday,4mice in each group were slau ghtered in day 3、7、11and 15d.The blood were collected and WBC, Gran, Lymph w ere tested before serum separation and hydroxyproline(Hyp) of IL-6,TGF-β determinati on. The histomorphology observation and immunohistochemical experiments of the wo unded skin were carried at the same time. The data obtained from the experiments w ere performed using the statistical software SPSS19.0,only those data that got a P<0. 05 were statistically significant.Results:1.The protein molecular weight of EADEC mainly concentrated between 15-35KD,45-55KD and 60-70KD;its inhibition zone of Escherichia coli, Salmonella,Str eptococcus,Bacillus subtilis were in the range of 6-10mm.2.EADEC could heal the wound on mice within 14 days,1 day earlier than the Jingwanhong group and 2.5 days earlier than positive control group.Mice in EADECg roup and Jingwanhong group could get a 100% healing rate in the wound,3% highert han those in the positive control group,the level of WBC, PLT,Gran in the bloodof t he mice in these 2 groups increased also.Compared with the positive control group, t he EADEC group and Jingwanhong group got more fibroblasts and collagen fibers, a nd these fibroblasts and fibers with abundant cytoplasm were all arranged in neat ro ws.The immature granulation tissue of the mice in these 2 group generate rapidlywith capillary in vigorous growth, significantly different with those in positive controlgroup (P<0.05).The content of Hyp,IL-6 and TGF-beta in EADEC group is better than the other two groups.Conclusion:1.An obvious function of promoting wound healing in local wound o n mice by treated with EADEC spray was observed, it could shorten wound healing time, reduce the injury of inflammation and improve the quality of healing.2.The mechanism of EADEC promoting wound healing are mainly by acceleratin g Hyp, TGF-beta secretion,and increase the synthesis of collagen, promoting blood c apillary and fibroblast proliferation;at the same time,it could accelerate the removal of necrotic tissue and foreign body by speeding up the generation of IL-6, blood WBC, PLT and Gran, thus enhance immunity, relieve the inflammatory reaction of mechanic al injury, reduce the risk of infection, and finally promote wound healing.