Preliminary Analysis On Active Component Of Coptidis Rhizoma Decoction Resisting Lipopolysaccharides Of Riemerella Anatipestifer

Riemerella anatipestifer is a Gram-negative short bacillus that can cause Riemerella Anatipestiferdisease. Many poultry is sensitive to this disease that is a huge threaten to aquaculture. The serotype ofRA is numerous. The immunization effect to RA is poor because among serotypes lack ofcross-protection capabilities. Antibiotic drugs have effect to this disease, but poultry resistance toantibiotic. Traditional Chinese medicine have good effect to resist RA. The main causative factor of RAis RA lipopolysaccharide(LPS). Previous studies found Coptidis Rhizoma Decoction made of Coptischinensis, Phellodendron amurense and Glycyrrhiza uralensis have goog results in against RA in vivoand vitro. With chemical solvent, Coptidis Rhizoma Dicoction were separated into different chemicalfractions. And compare the anti-endotoxin activity of three different fractions isolated from CoptidisRhizoma Dicoction.The Coptidis Rhizoma Dicoction is made by mixing Coptis chinensis, Phellodendron amurense andGlycyrrhiza uralensis according to the ratio of1:1:2, with water extraction and alcohol sedimentationmethod. The concentration of the drug is174.0g/L. Multiple diluting this drug into Ethanol Extracts ofCoptidis Rhizoma Decoctin of10concentration gradient,87×20、87×2-1、87×2-2、87×2-3、87×2-4、87×2-5、87×2-6、85.7×2-7、87×2-8、87×2-9g/L.Choose disc diffusion test and tube broth assays to study the effect of EeCrd against RA invitro.Every disc contain200μL EeCrd. The result of disc diffusion test showed that the group of88×2-5g/L have no ingibition zone, the group of88×20g/L have the biggest ingibition zone. The result of tubebroth assays showed that the group of87×2-1g/L have the least bacteria in the tube. EeCrd havesignificant effect in against RA in vitro.The LPS is extracted from serotype2Riemerella anatipestifer by hot phenol water method.200μLdifferent concentration of EeCrd and10μL LPS interact in constant temperature incubator, in37℃,200r/min, incubating11h. Quantitatice detection of the LPS of the testing samples with chromogenictechnique. Results showed that with the iecreased of the EeCrd’s concentration, LPS is damaged moreand more. The group of87×2-1g/L destroyed almost all LPS.With microwave-assisted extraction, the EeCrd was devided into three fractions, n-hexane extract(NHE), ethyl acetate extract(EAE) and n-butanol extract(NBE). This three fractions was dilutedinto1,0.5,0.25mg/mL.200μL different extract of different concentration, and10μL LPS interact inconstant temperature incubator, in37℃,200r/min, incubating11h. Quantitatice Detection of LPS oferery sample with chromogenic technique. Results showed that EAE has a significant role in theelimination of LPS in vitro.Some bioactive components in this three extract fraction was detected with thin layerchromatograph(TLC). Results showed that EAT contains a lot of flavonoids and little alkaloids andanthraquinones.