| Hepatitis B is one kind of infective disease which caused by HBV and difficult to be cured. The morbidity rate is very high in china. Hepatitis B could lead to cirrhosis and liver cancer. The disease is harmful to human beings. It is an urgent task for us to look for both safe and efficacious anti-HBV drugs. The high efficiency anti-HBV drugs-screening system both in vitro and in vivo have been established and applied. The contents of this article are as follows: 1. Nine kinds of nature durgs were extracted and tested by HepG2.2.15 cells the anti-HBV drugs screening system in vitro. The results show that extraction from the Semen Litchi has strong function in anti-HBV in vitro. 2. Cytotoxicity was determined by SRB(suforhodamine B Protein dye-binding)assay and the content of HBV-DNA was determined by polymerase chain reaction(PCR)method in the anti-HBV drugs screening system in vitro. 3. Throught PCR method, the Guilin brown spot ducklings congenitally infected by duck hepatitis B virus(DHBV)were Screened out, then the effect of durgs on DHBV in vivo was studied. 4. Optical density mensuration, cell culture and FQ-PCR technique were used to study the inhibitory effects of the extraction from the Semen Litchi on the expression of HBsAg, HBeAg and the content of HBV-DNA in HepG2.2.15 cells. In 96 holes plate test, i(t800,400,200,100 μg/mL)had evident action to inhibition on expression of HBsAg and HBeAg in HepG2.2.15 cells on the thirth and sixth experiment day,compared with contral group(P<0.01). In 24 holes plate test, it(400, 200, 100, 50 μg/mL)had evident action to inhibitiong on expression of HBsAg and HBeAg in HepG2.2.15 cells on the thirth, sixth and ninth experment day,compared with contral group(P<0.01). FQ-PCR showed that the extraction(400 μg/mL)turned HBV-DNA in HepG2.2.15 cells medium negative.The exraction from the core of Semen Litchi has strong function in anti-HBV in vitro. 5. The anti-HBV effect of extraction from the core of Semen Litchi. By industrial produced way was also investigated both in vitro and in vivo examination. 6. Inhibitory effects on expression of HBsAg, HBeAg in HepG2.2.15 cells with the industrial extraction from the core of Semen Litchi were stdudied. The extraction(800,400,200,100 μg/mL)had evident action to inhibition on expression of HBsAg and HBeAg in HepG2.2.15 cells on the thirth and sixth experiment day,compared with contral group(P<0.01).Inhibition rate to HBsAg of industrial extraction(400 μg/mL)was 73.00 % and 94.00 % in 3 day and in 6 day. Inhbition rate to HBeAg was 49.00 % and 62.00 % in 3 day and in 6 day. The result showed that the industrial extraction from the core of Semen Litchi has also obvious function to anti-HBV in vitro. 7. Guilin brown spot ducklings congenitally infected with DHBV was used as animal modal to study the anti-HBV action of industrial extraction from the core of Semen Litchi. After medicated with the industrial extraction(2 g/kg/day)5 day, 10 day ang 15 day. DHBV-DNA in ducks sera were detected with Optic density valus(OD value)DHBV-DNA in ducks sera of the industrial extraction treatment group was obviously lower than before treatment. There was significant difference(compared with virus control group P<0.05, compared with before treatment P<0.01), The inhibition rate was 62.5 % in after medicated with the industrial extraction 5 day, The result showed the industrial extraction(2 g/kg/day)can inhibit the replication of DHBV-DNA in GuiLin brown spot ducklings. GuiLin brown spot ducklings of DHBV-modal were killed to observe pathological in liver tissue. The resuts showed that industrial extraction(2 g/kg/day)has obvious action to protest livers(compared with control group P<0.05). |
