Studies On The Anti-biotic, Anti-tumor And Immunology Enhancement Activities Of Earthworm Tissue Extracts

Earthworms,found all over the country, and have effects on many diseases,it's one of the most precious traditional chinese medicine. Researchers have separated many active materials from the earthworms, as antibacterial peptides. But, there have not been reported in the literature that studies on the anti-biotic, anti-tumor and immunology enhancement activities of earthworm tissue extracts.Objective: Separated biological activity materials from earthworm, research it antibiotic, anti-tumor and immune enhancing activities,explore its mechanisms.Our results can provide the basis for further research of antibiotic drugs and anti-tumor drugs.Methods: Preparation earthworm crude extract by the methods of fractional precipitation. Estimate molecular weight of the earthworm crude extract by SDS-PAGE and Tricine-glycerol SDS-PAGE.Research the earthworm crude extract anti-bacterial activity by plate punch method and select the strong activity crude extract. Minimal inhibitory concentration (MIC) was determined by microamount plate method. Coarse isolate the strong activity crude extract by glucosan G- 15,collect the essential component as crude separation extract. Analysis the crude separation extract by high performance liquide chromatography(HPLC).Determin enhancement of the crude separation extrat on splenic lymphocytes by MTT, and determine the inhibitory in vitro on S-180 sarcoma cell growth by the same methods.To research the mechanism of anti-tumor, we established S-180 sarcoma tumor models in mice,the mice were divided by the different dose of the crude separation extract. Sixty kunming mice were randomly allocated to an six group. There are ten mice in each group.(1)Not vaccination S-180 group:the mice were perfused orally into stomach with 0.2mL nomal saline per every day and not transplanted tumor cell;(2)S-180 model control group:the mice were transplanted tumor cell and perfused orally into stomach with 0.2mL nomal saline per every day;(3)Crude separated extract groupâ… :the mice were transplanted tumor cell and perfused orally into stomach with 100mg/kg crude separated extract per every day;(4) Crude separated extract groupâ…¡:the mice were transplanted tumor cell and perfused orally into stomach with 300mg/kg crude separated extract per every day:(5) Crude separated extract groupâ…¢:the mice were transplanted tumor cell and perfused orally into stomach with 500mg/kg crude separated extract per every day;(6)S-180 Cy control group:the mice were transplanted tumor cell perfused orally into stomach with Cy, 20mg/kg per every day.Determine effects of crude separation extract on macrophage,tumor inhibition rate,thymus index,spleen index. All of these were compared with the group of anticancer drugs Cy.Results:Separated three crude extracts from earthworm,and the 9 times acetone crude extract had the best inhibiting effect on E.coli, Staphylococcus aureus and proteus bacilli,and the diameter of the inhibition areas is 1.7cm, 1.3cm, 1.5cm.The MIC to E.coli is 25mg/L,and the MIC to Staphylococcus aureus and proteus bacilli are both 50mg/L.The crude separated extract was separated from 9 times acetone crude extract, and the molecular weight was 3313Da-7823 Da.The crude separated extract have much enhancement on splenic lymphocytes,and can inhibit S-180 sarcoma cell growth in vitro and in vivo.The crude separated extract also can enhance the thymus index and spleen index,enhance the phagocytic activity of macrophage, and greatly improve the immunity of S-180 tumor bearing mice.