Guangxi vine tea extract TTF anti-duck hepatitis B virus in
【Abstract】 Objective To observe the tea from the Guangxi rattan extracted an effective part of TTF role of anti-duck
hepatitis B virus. Elections of one-day-old ducklings Beijing,
artificial infection of duck hepatitis B virus (DHBV) animal model, the application of the dot-blot hybridization observed
TTF duck serum duck hepatitis B virus DNA (DHBV-DNA) effects.
Results DHBV infected ducks in the first 7 days after
infection TTF oral medication, 75 ~ 150 mg / kg group 2 times / d, 10 d for duck serum DHBV-DNA levels have a certain
inhibitory effect; 300 mg / kg group, infected duck serum DHBV-DNA level of inhibition significantly, stopping 3 d no significant anti-jump phenomenon. Conclusions TTF could
significantly inhibit the DHBV-DNA replication, suggesting
that TTF has anti-HBV effect.
Key words vine tea duck hepatitis B virus (DHBV) animal model of duck hepatitis B
Abstract: ObjectiveTo investigate the anti-HBV effect of
TTF.Methods1-day old Beijing ducklings were used as the hepatitis B animal model. DHBV DNA was detected by dot-blot
hybridization test. ResultsAfter medicated with TTF for 5 days and 10 days, optical density value (OD value) of DHBV-DNA in
ducklings sera was obviously lower than that before treatment, there were different influences in different groups and rebound phenomenon didn `t appear in both TTF-groups 3 days
after stopping medication.ConclusionTTF is efficient in
treating DHBV, which indicates that TTF has an anti-HBV
Key words: Teng cha; Duck hepatitis B virus; Duck hepatitis Model
Vine Tea Department Vitaceae Ampelopsis species Ampelopsis Ampelopsis grossedentata (Hand-Mazz) WTWang of the
Spear leaves, in Guangxi, Guangdong, Yunnan, Guizhou and Fujian are the distribution, also known as sweet tea vine, Tian Po Tea, Wu Lian, red cairica big tooth snake wine. Its sweet, light, sexual cool, with heat-clearing and detoxifying,
rheumatism, strong bones and muscles, swelling of the
effectiveness of pain. Vine tea folklore has been around for hundreds of years of history, is the home of the Yao people standing medication. People used to treat jaundice hepatitis, cold fever, sore throat, acute conjunctivitis, or water to
cook outside the washing treatment of carbuncle sore poisoning caused by damp, etc. [1 ~ 3]. Vine is the main component of tea flavonoids, the content is about 45.52% . Cheng  The results show that tea flavonoids on 2215 Vine cell inhibition
rate of HBsAg secreted by 43.14%, half of the effective dose (IC50) of 40.21 μg / ml, therapeutic index (SI) of 3.11; to 2215 cells in the secretion of HBeAg inhibition rate of 33.76%; pairs of HBV-DNA have a certain inhibitory effect. To this end, the author of the TTF in vivo anti-duck hepatitis B
virus in experiments. Are reported below.
1.1 Subjects of drug TTF is light yellow crystalline powder, Chinese medicine chemistry teaching and research provided by the school, the content over 90%.
1.2 Virus duck hepatitis B virus DNA (DHBV-DNA) strongly
positive sera were collected from Shanghai duck, -70 ? to
1.3 reagent DMSO (AR, Guangzhou Xingang Chemical Co., Ltd.); α-32P-Dctp (Beijing Furui Biological Engineering Company); nick translation kit (Puluomaige Corporation (Promega Co.); Sephadex G-50 ( Ficoll); PVP (Sweden Pharmacia
Corporation); SDS (West Germany Merck Company); fish sperm DNA, bovine serum albumin were purchased from Bio-Physics,
Chinese Academy of Sciences; nitrocellulose membrane 0.45 μm
1.4 Instrument BP211D ten thousandth g electronic balance (Sartorius); microplate reader (Austria SUNRISE); trace plus sampler (France Gilson); Biofuge Stratos Eppendorf (Germany Heraeus); Block automatic electric pressure steam sterilizer (Shanghai SHEN instrumentarija).
1.5 positive control drug lamivudine (3TC), Glaxo Wellcome pharmaceutical company, B008923.
1.6-day-old animals, a Beijing duck, purchased from Beijing ahead duck farm animals.
2 methods and results
2.1 TTF anti-duck hepatitis B virus experiment
2.1.1 duck hepatitis B virus infection in 1-day-old
Beijing duck, after intravenous injection of Shanghai duck leg tibia DHBV-DNA positive duck serum, 0.2 ml / only at 7 d after infection, blood, separation of serum, -70 ? to save for
2.1.2 Drug treatment of experimental DHBV infection in ducklings 7 d after drug treatment trials were randomly divided into groups, each group 6, administration of component 3-dose group, respectively, 75,150,300 mg / kg group, oral, 2
times / d, continuous administration of 10 d. Of viral control group (DHBV), with saline instead of drugs. Positive medicinal 3TC, oral administration of 50 mg / kg, 2 times / d, continuous administration of 10 d. In the first 7 days after infection that before treatment (T0), medication 5 days (T5), medication 10 days (T10) and 3 days after withdrawal (P3), posterior tibial vein taken from the leg of the duck blood, separation of serum, -70 ? to save for inspection. Reposted
elsewhere in the paper for free download http://
Take the above question to check duck serum, each batch of films at the same time points were determined duck serum DHBV-DNA level of dynamic. Nick translation kit according to manual methods, using 32P marked DHBV-DNA probe, and to make
duck serum dot blot hybridization, autoradiography diaphragm
spots, in the ELISA test to measure the OD value (filter of 490 nm), calculated serum DHBV-DNA density to dot
hybridization samples OD value as the value of DHBV-DNA
levels. Each duck will be at different times after treatment (T5, T10), and 3 days after withdrawal (P3) of serum DHBV-DNA
levels and the same group before administration (T0) OD value, using paired t test, analysis of variance significantly determine the drugs on the inhibitory effect of virus
infection. According to a formula in each group at different times after administration ducks (T5, T10), and withdrawal on day 3 (P3) of serum DHBV-DNA inhibition rate%, and mapping,
comparing each group duck serum DHBV-DNA inhibition rate of
the dynamics of .
DNA inhibitory rate (%) = before administration (T0) OD value - after the administration (T5, T10, P3) OD values before administration (T0) OD value × 100%
The administration of the treatment group at different
times of DHBV-DNA inhibition rates were the same time with the virus control group, comparing the rate of DHBV-DNA
inhibition, using grouped t test for statistical analysis, computing t, P value, analysis of significant differences to determine efficacy.
2.2.1 1-day-old Beijing duck infected with DHBV, serum DHBV-DNA dynamics ducks infected with DHBV-DNA after oral
administration of saline DHBV-DNA dot blot hybridization
results shown in Figure 1. 30 ducks experimentally infected
with serum DHBV-DNA all were positive. Virus control group, six ducklings the first 7 days after infection, serum DHBV-DNA
positive for all the experimental full within 21 d of serum DHBV-DNA levels after infection basically stable. Whole different time fluctuations of serum DHBV-DNA, infection and
post-treatment 10 d saline serum levels of DHBA-DNA was no
2.2.2 of the DHBV infected duck serum DHBV-DNA results
show that the impact of, TTF low-dose group 10 days after
stopping administration of the first 3 d with the administration before (T0) Comparison of calculated inhibition rate, matching analysis and comparison, a significant and very