Guangxi vine tea extract TTF anti-duck hepatitis B virus in experimental study_1016

By Daniel Gardner,2014-10-30 18:57
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Guangxi vine tea extract TTF anti-duck hepatitis B virus in experimental study_1016

Guangxi vine tea extract TTF anti-duck hepatitis B virus in

    experimental study

     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% [4]. Cheng [5] 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 Materials

     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

    (Amersham Corporation).

     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://

     2.1.3 Detection

     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 Results

     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

    significant difference.

     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

significant (P

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