Dissociates And Pepurates The 3-30KD Antibiotic Peptide Of Earthworm And Studies On The Antibacterial Effect

Antibacterial peptides(APBs) have the favourable bacteriostasis effective and won't bring the drug fast reaction in living organism, so there are more and more research on mechanism of action, physico-chemical property and so on.. It is the destination of study that the 3-30KD APBs of earthworm was abstracted and depurated, then to analyze the bacteriostasis effective at out-of-body and ntracorporal.After daping2 earthworm was cleaning, induction by lightning stroke, ultrafiltrating by 3,30KD ultrafiltration centrifuge tube, antibacterial peptides was isolated from earthworm by Sephadex G-50 gel filtration chromatoraphy and ultrafiltration and SP-Sepharose Fast Flow ion exchang chromatoraphy, getting two wave crest of protein named H1,H2.Their molecular mass are between 5.856-7.823KD and about 20.10KD by using Tricine-SDS-Page.Using microdetermination assay bacteriostasis activity of H1, H2, the experiment result show that H1, H2 have the different level wih the restrain and killing effect to colibacillus, Staphylococcus aureus, paratyphosus bacillus, cerea bacillus, pastometer bacillus, streptococcus but bacillus and Bacillus subtilis, and that APBs of earthworm has broad-spectrum ntisepticize keeping beneficial bacterium. After using H1, H2 dealing with pastometer bacillus nutrient solution, under the scanning electron microscope observation result show that H, H2 have a different eaffect for pastometer bacillus. They can destroy cell envelope and cell wall of pastometer bacillus, and make their cage unclear, intracellular fluid leaking out, vacuolus to bring about in thallus, even fragmentation death, so it revealed initial bacteriostasis mechanism on APBs of earthworm.In vivo experiment is treating pastometer bacillosis with the earthworm extract. Through assay WBC and NC, the development bacterium in vivo, the value of TNF-a and NO, the content mRNA about TLR4, the result show that APBs of earthworm can stimulate immunereaction, enhance leukocytotic hyperplasy at catadrome pathopoiesis, restrain bacterium growth in vivo also kill them, contain the inflammatory factor TNF-a, NO increasing fast, decrease the content of TLR4 mRMA induced by LPS, palliate inflammatory damage of bellows, so APBs of earthworm can treat this sick in force.