Effect Of Phyliathus Complex Combining With ADV To Treat LRHB And Antiviral Immunologic Mechanism Research

For many years of experiment and clinical research, repeatedly selection and continuous optimization, our study use Phyliathu urinaria Linn (PUL), Astragalus membranaceus, pseudo-ginseng, liquorice, etc.to compose the capsule of Phyliathus Complex (PC).We hope to explore a new generation of traditional Chinese herbal combination to treat chronic hepatitis B.Before the clinic study, we have accomplished a series of experiment research of anti-HBV in vitro,and an experiment research about the intervention of HBV transgenic mouse immunopathogenesis model.We proved the efficiency of treatment.In the research of anti-HBV in vitro,we choose2.2.15cell as a vitro model,HBsAg, HbeAg as drug effect of the preliminary screening targets.Though MTT method measure cell toxicity concentration of medicine,and calculate therapeutic index to evaluate medicine efficiency. Results show that, the therapeutic index of Phyliathus Complex inhibiting HBsAg in2.2.15cell is8.4.So Phyliathus Complex has high-efficiency and low-toxin to inhibit HBsAg and HBeAg in vitro.The rate of HBsAg negative-inverted with using Phyliathus Complex of medium-dose, high-dose and ADV in Tg mouse are compared respectively at10d,21d, after without drug3d. There is no significant difference.ADV has an effect of reducing serum HBV-DNA load in HBV Tg mouse.But after without drug3d, it has a bounce.Phyliathus Complex of medium-dose, high-dose has an effect of reducing serum HBV-DNA load in HBV Tg mouse. Specially, high-dose has a significant effect. HBV-DNA load can't reduce to the level of ADV group, but there is a significant difference before and after treatment, and don't appear bounce after without drug. We can see that Phyliathus Complex have a stable and durable effect. In model group,Tg mouse liver pathological changes is similar to the expression of clinical mild hepatitis, with increasing the dose, inflammation can be relieved. The hepatic plates of high-dose group arrange appropriately, there is no fibrous tissue hyperplasia, but perivascular inflammatory infiltrates mildly. Phyliathus Complex of medium-dose, high-dose and ADV all can decrease HBcAg expression in liver issue, especially in high-dose group.Phyliathus Complex can up regulate CD3+,CD4+,CD4+/CD8+, and down regulate level of CD8+T cell, has a positive immune regulation for T cell lymphocyte, specially in high-dose group.ADV also can improve immune function of HBV transgenosis mouse cell (such as percentage of CD3+), but the effect of ADV is lower than Phyliathus Complex.Using Phyliathus Complex of high, medium,small dose and ADV can all increase serum IL-2and TNF-r content in HBV transgenosis mouse,there is a significant difference compared with control group.And after using Phyliathus Complex of high-dose and medium-dose, IL-2and TNF-γ content is higher than normal group.In Phyliathus Complex of medium-dose group, CD3+,TNF-γ have a positive correlation with reduction of HBV-DNA load before and after treatment.In high-dose group, changes of CD4+,CD8+,CD4+/CD8+have a positive correlation with reduction of HBV-DNA load before and after treatment.Phyliathus Complex combining with ADV treat Lamivudine resistant hepatitis B (LRHB) for52weeks.ALT recovery rate, the rate of HBV-DNA and HBeAg negative-inverted are all higher than single AVD group, and there is a significant difference (P<0.05or P<0.01).After52weeks treatment, the complete response rate of Phyliathus Complex combining with ADV is25%,remarkable higher than ADV group(10%), X2=9.07,P<0.01.Using Phyliathus Complex combining with ADV group after52weeks treatment, CD4+, CD3+percentage and NK cell of patients all increase, but CD8+percentage of two group all decrease, and has a positive correlation with reduction of HBV-DNA load. HBV specific type CDg+T cells destroy infective liver cell, eliminate HBV in liver, thus reduce amount of HBV in cells.It shows, Phyliathus Complex combining with ADV can improve abnormal situation of T cell lymphocyte in LRHB patients, and increase NK cell activity, enhance antiviral efficiency. Meanwhile, Phyliathus Complex combining with ADV also can regulate immune function; make low or abnormal immune to normal situation,so it shows a better anti-HBV effect.