Screening Of Anti-duck Viral Hepatitis Chinese Herbal Compound Prescription And Its Machanisam

Duck viral hepatitis(DVH)is a kind of rapidly spreading,infectious and fatal infectious disease with high morbidity and high mortality caused by duck hepatitis A virus(DHAV).Once the disease occurred,a huge loss appeared.The experiment firstly screened out compound that can inhibit DHAV.Secondly,prescription was formed and the anti-DHAV activities of optimum were tested in vivo and in vitro.Pharmacodynamical and pharmacological effects of optimized formula were studied to hunt for prescription that has good effect for clinical prevention and control of DVH.Experiment ?:The screening of the anti-DHAV Chinese herbal compound prescription In order to develop a new anti-Duck viral hepatitis(DVH)Chinese herbal compound prescription,this paper had filtrated 8 compounds effective to DVH from epimedium flavone(EF),,notoginseng flavone(NF),wild chrysanthemum flavone(WCF),baicalin(BA),geniposide(GP),rehmannioside(RN),Radix Rehmaniae Preparate Polysaccharide(RRPP)and selfheal flavonoids(SF)to form prescriptions.The prescriptions were experimented in vitro and in vivo for anti-duck hepatitis virus(DHV)studies to find ones that were effective to DHV both in vitro and in vivo.The results suggested that:all compoounds had anti-DHAV activities(NF>GP>BA>EF>RN>WCF>SF>RRRP).Prescriptions of EF-NF-WCF-BA,EF-RRPP-WCF-BA,EF-RN-WCF-BA,EF-RRPP-RN-WCF-BA were effective in vitro while EF-RRPP-WCF-BA and EF-RN-WCF-BA were also effective in vivo to DVH.Considering factors of economic and drug resource,prescription of EF-RRPP-WCF-BA is chosen as the optimal one to anti-DVH.Experiment ?:The therapeutical effect of EF-RRRP-WCF-BA to DVH The purpose of this assay is to ensure the therapeutical effect of optimal on DVH.135 ducklings were randomly divided into 3 groups(EF-RRRP-WCF-BA,VC and BC group).The ducklings of EF-RRRP-WCF-BA and VC group were intramscular injected with 5 LD50 DHAV while duklings of BC group were intramscular injected with same volume physiological saline.One hour after DHAV attack,ducklings of EF-RRRP-WCF-BA group immediately drinking water mixed with drug(3 mg/feather)with one time per day in consecutive 5 days.The blood sample and plasma sample of 5 feathers of each group were collected.Observed the dynamic death of ducklings and determined the level of the plasma hepatic injury evaluation indexes(ALP,AST,ALT,etc.),inflammatory cytokines(IL-6,IL-8,HGF)and blood DHAV relative expression quantity.The results showed that EF-RRRP-WCF-BA could inhibit the abnormal change of the evaluation of hepatic injury indexes,increase the level of HGF and IL-6 and decrease the relative expression of at 54 h.It indicated that EF-RRRP-WCF-BA had definite therapeutic effect to DVH.Experiment III:Protective effect of EF-RRPP-WCF-BA for oxidative damage induced by DHAV of ducklings In order to further study the anti-DHAV effect of EF-RRPP-WCF-BA,this experiment observed SOD,CAT,GSH-Px,NOS,MDA changes of ducklings plasma at 4 h,8 h,54 h after intramscular injection with 5 LD50 DHAV and duck embryo liver cell culture supernatant after infection with 50 TCID50 DHAV(animals grouping and treatment as experiment II).Results showed that EF-RRRP-WCF-BA could raise the level of SOD,CAT,GSH-Px in vitro and in vivo,decrease NOS and MDA levels in vitro and in vivo.The tendency of evaluation indexes in vitro and in vivo were the same in general.It can be included that EF-RRPP-WCF-BA can alleviate the oxidative damage caused by DHAV.Experiment IV:Immune regulation function of EF-RRRP-WCF-BA on ducklings To observe the immune effect of EF-RREP-WCF-BA on DVH,immune related cytokines including interleukin 2(IL-2),interleukin 4(IL-4)and interferon gamma(IFN-y)were detected in the experiment at 4 h,8 h,54 h after intramscular injection with 5 LD50 DHAV(animals grouping and treatment as experiment II).The effect of EF-RRRP-WCF-BA on lymphocyte proliferation was measured via lymphocyte transformation test.The results showed that EF-RRPP-WCF-BA can improve IL-2,IL-4 and IFN-y levels,demonstrating that it had immune enhancement effect.Meanwhile,lymphocyte transformation test showed that it can stimulate T lymphocyte proliferation.