The Study Of Anti-HBV And Anti-immune Liver Injury Action And Mechanism Of Compound Of PHyllanthus Urinaria Ⅱ

ObjectiveTo evaluate the anti-HBV effect of the compound of PHyllanthus Urinariaâ…¡(CPUâ…¡) decoction.We use the 2.2.15 cell line as cell model and congenitally DHBV positive ducks as animal model.In the experimental study of the anti-immune liver injury In vivo,animal model of acute liver injury in mice included by concanavalin A was used for studying the therapeutic effects of CPUâ…¡,and providing scientific gist to the widely use of CPUâ…¡in the clinic.Experimental contents1.The experimental study of anti-HBV effect of CPUâ…¡1.1 In-vitro experimental study of the anti-HBV effect of CPUâ…¡The 2.2.15 cell line was used as cell model.CPUâ…¡was diluted into eleven dosage groups.The HBsAg and HBeAg in the supernatant were tested by ELISA assay.The cytotoxic of drug to the cell was evaluated by the MTT assay.The results showed that the TC₅₀ of CPUâ…¡to 2.2.15 cell line was 16.94 mg/ml.The IC₅₀ of CPUâ…¡to HBsAg,HBeAg was 7.82mg/ml and 7.60mg/ml respectively,the TI of CPUâ…¡was 2.17 and 2.23.The results showed that CPUâ…¡can inhibit virus replicantion on the 2.2.15 cell in-vitro.1.2 In-vivo experimental study of the anti-DHBV effect of CPUâ…¡In-vivo,ducks with hepatitis B virus was used as animal model.The serum was collected by the time points of T0,T7,T14,T21,T28 and P5 in accordance with the time point of drug intragastric administration.The quantity of DHBV in these specimens was evaluated by quantitative PCR method.The results that the serum DHBV load of both the high and middle dosage of CPUâ…¡at T7,T14,T21,T28 and P5 were obviously lower than that at T0(P <0.05,0.01);and obviously lower to that of model group(P<0.05,0.01). The experiment indicated CPUâ…¡could inhibit virus replicantion rapidly.There was no rebound phenomenon appearing in CPUâ…¡after stopping medication.2.Immunity experimental study of the anti-liver injury effect of CPUâ…¡2.1 The effect of CPUâ…¡on the activity of serum ALT and IL-6 that from the acute immune liver injury in miceIn the experimental study of anti-immune liver injury,animal model of acute liver injury in mice induced by concanavalin A was used for studying the therapeutic effects of CPU II with high and low dosage groups(33.6g/kg,16.8g/kg),bifendate as the positive control.Activities of serum alaninc transaminase(ALT),IL-6 in liver tissue was determined, and hepatic histopathological change were observed.The results showed that ALT activity in the high and low group of CPUâ…¡were obviously lower than that of model group(P<0.01),and the ALT activity in the high group of CPUâ…¡was obviously lower than that of the low group and bifendate group,that were significant difference(P<0.01).The activity of IL-6 was lower than that of model group,and that was significant difference (P<0.05).The hepatic histopathological change were alleviated in both CPUâ…¡with high and low dosage group.The experiment indicated CPUâ…¡has a good protective effect on liver injury in mice induced by Con A by suppressing the activation of the T-lymphocytes and reducing the release of IL-6.2.2 The effect of CPUâ…¡on the activity of serum TNF-a that from the acute liver injury in miceIn the experimental study of anti-immune liver injury,animal model of acute liver injury in mice included by concanavalin A was used for studying the therapeutic effects of CPUâ…¡with high and low dosage groups (33.6g/kg,16.8g/kg),cyclophosphamide as the positive control.Activities of serum tumor necrosis factor-a(TNF-α) was determined.The results showed that The TNF-αof high and low dosage group of CPUâ…¡was obviously lower than that of model group(P<0.01,0.05),and the TNF-αof high dosage group was obviously lower than that of low dosage group,that was significant difference.The experiment indicated CPUâ…¡has a good protective effect on liver injury in mice induced by Con A by reducing the release of TNF-α. 1.There are good effect of CPUâ…¡on anti-HBV in vitro and in vivo2.There are good effect of CPUâ…¡on anti-immune liver injury that induced by Con A by suppressing the activation of the T-lymphocytes and reducig the release of TNF-αand IL-6.