Therapeutic Effects Of Aylsl And Aylsr On DHBV-Induced Chronic Hepatitis In Duck

Objective:To establish the chronic hepatitis B duck Model and study the therapeutic effects of AYLSL and AYLSR on DHBV-induced chronic hepatitis in duck.Methods:One-day old Guangxi shelducks were infected with 0.2mL Duck hepatitis B virus (DHBV). The infected ducks, detected by PCR 52 days after infection, were divided into model group, AYLSR high-dose and low-dose group (20, 10g.kg-1), AYLSL high-dose and low-dose group (20,10g.kg-1), and the uninfected ducks were served as normal control group. Animals are bred under the same conditions. The corresponding drugs were successively administrated for 150 days after 60 days of DHBV infection. Normal control group and model group were intragastric administrated with the same volume of double steaming water. Serum samples were colleted at 0th (T0),30th (T30),60th (T60),90th (T90), 120th (T120), and 150th (T150) day of treatment, and withdrawal at 7th (P7) day. Each duckling was dissected to obtain the liver immediately with pentobarbital sodium anesthesia, after the last time collecting blood sample. Blood and liver samples were appropriate processed and used for the detection of relevant indicators.(1)The level of DHBV-DNA was detected with FQ-PCR, the content of DHBsAg and DHBeAg were measured using ELISA kits and the activity of ALT and AST were detected using fully automatic biochemical analysis system.(2)The activity of superoxide dismutase (SOD) and glutathion peroxidase (GSH-Px), and the level of malondialdehyde (MDA) and glutathion (GSH) in liver homogenate were determinated, using corresponding kits. Hematoxylin-eosin (HE) staining was used to observe the damage degree of liver cells.(3)The DNA of hepatic tissue was extracted, which were used to determinate the content of DHBV-cccDNA in each group, using FQ-PCR.(4)ELISA kits were used to measure the concentrations of Col-1, LN, and HA in serum, the content of hydroxyproline (Hyp) in liver was detected using corresponding kit.(5)The total RNA of hepatic tissues were extracted, and RT-PCR was used to evaluate the mRNA expression of Col-1 and TIMP-2.Results:(1)Compared with model group, AYLSL groups (20, 10g.Jkg-1) and AYLSR groups (20, 10g.kg-1), can significantly reduce the content of DHBV-DNA, DHBsAg and DHBeAg, and reduce the activity of ALT and AST in serum (P<0.05 or P<0.01)(2)Compared with model group, AYLSL groups (20, lOg.kg-1) and AYLSR groups (20, lOg.kg-1), can significantly increase the activity of superoxide dismutase (SOD) and glutathion peroxidase (GSH-Px), increase the level of glutathion (GSH) (P<0.05 or P<0.01) Hematoxylin-eosin staining showed that AYLSR and AYLSL can significantly decrease the damage degree of liver cells.(3)Compared with model group, AYLSL group (20g.kg-1) and AYLSR group (20g.kg-2) can significantly decrease the content of DHBV-cccDNA in duck hepatic tissues (P<0.05 or P<0.01)(4)Compared with model group, AYLSL groups (20,10g.kg-1) and AYLSR groups (20, 10g.kg-1), can significantly decrease concentrations of Col-1, LN, and HA in serum, and decrease the content of malondialdehyde (MDA) in liver tissue.(5)Compared with model group, AYLSL groups (20, 10g.kg-1) and AYLSR groups (20, 10g.kg-1), can significantly the mRNA expression of Col-I and TIMP-2 in hepatic tissues (P<0.05 or P<0.01)Conclusions:AYLSR and AYLSL have certain therapeutic effect on chronic hepatitis caused by DHBV, and shown the inhibition against deterioration of liver disease. The mechanism may relate to antiviral, inhibition of lipid peroxidation and inhibition of liver fibrosis.