RESUMO
BACKGROUND: Hepatitis B virus (HBV) infection remains a major global health burden, due to the increasing risk of complications, such as cirrhosis and hepatocellular carcinoma. Novel anti-HBV agents are critical required. Our previous study suggested that Artemisia argyi essential oil (AAEO) significantly inhibited the replication of HBV DNA and especially the secretion of hepatitis B antigen in vitro. PURPOSE: The aim of this study was to prepare AAEO loaded nanostructured lipid carriers (AAEO-NLCs) for the delivery of AAEO to the liver, investigated the therapeutic benefits of AAEO-NLCs against HBV in a duck HBV (DHBV) model and explored its potential mechanism. STUDY DESIGN AND METHODS: AAEO-NLCs were prepared by hot homogenization and ultrasonication method. The DHBV-infected ducks were treated with AAEO (4 mg/kg), AAEO-NLCs (0.8, 4, and 20 mg/kg of AAEO), and lamivudine (20 mg/kg) for 15 days. The DHBV DNA levels in the serum and liver were measured by quantitative Real-Time PCR. Pharmacokinetics and liver distribution were performed in rats after oral administration of AAEO-NLCs and AAEO suspension. The potential antiviral mechanism and active compounds of AAEO were investigated by network pharmacology and molecular docking. RESULTS: AAEO-NLCs markedly inhibited the replication of DHBV DNA in a dose-dependent manner and displayed a low virologic rebound following withdrawal the treatment in DHBV-infected ducks. Moreover, AAEO-NLCs led to a more pronounced reduction in viral DNA levels than AAEO suspension. Further investigations of pharmacokinetics and liver distribution in rats confirmed that NLCs improved the oral bioavailability and increased the liver exposure of AAEO. The potential mechanisms of AAEO against HBV explored by network pharmacology were associated with signaling pathways related to immune response, such as tumor necrosis factor, nuclear factor kappa B, and sphingolipid signaling pathways. Furthermore, a total of 16 potential targets were obtained, including prostaglandin-endoperoxide synthase-2 (PTGS2), caspase-3, progesterone receptor, etc. Compound-target docking results confirmed that four active compounds of AAEO had strong binding interactions with the active sites of PTGS2. CONCLUSIONS: AAEO-NLCs displayed potent anti-HBV activity with improved oral bioavailability and liver exposure of AAEO. Thus, it may be a potential therapeutic strategy for the treatment of HBV infection.
Assuntos
Artemisia , Vírus da Hepatite B do Pato , Neoplasias Hepáticas , Óleos Voláteis , Ratos , Animais , Simulação de Acoplamento Molecular , Óleos Voláteis/farmacologia , Farmacologia em Rede , Ciclo-Oxigenase 2 , Antivirais/farmacologia , Vírus da Hepatite B/genética , Vírus da Hepatite B do Pato/genéticaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Formula Le-Cao-Shi (LCS) is a traditional Chinese medicine (TCM), which has long been used as a folk remedy against hepatitis B in China. The present study was conducted to evaluate the anti-hepatitis B effects of aqueous extract of LCS in vivo and in vitro. MATERIALS AND METHOD: we investigated the anti-HBV effects of LCS in vivo and in vitro with duck hepatitis B model and HepG2.2.15â¯cell line model, respectively. The serologic and cellular biomarkers and the histopathological changes were examined. RESULTS: By a duck hepatitis B model, the extract of LCS was found to restrain the expressions of duck hepatitis B surface antigen (DHBsAg), hepatitis B e antigen (DHBeAg), and HBV-DNA (DHBV-DNA). Moreover, LCS could decrease the levels of aspartate and alanine aminotransferases (AST and ALT) and ameliorate duck liver histological lesions. Correspondingly, in a HepG2.2.15 cellular model, LCS could also significantly inhibit the secretions of HBsAg and HBeAg. CONCLUSION: LCS exerted potent anti-hepatitis effects against the infection of HBV. The above results demonstrated the first-hand experimental evidences for the anti-hepatitis B efficiency of LCS. Our study provides a basis for further exploration and development of this promising compound prescription to treat hepatitis B disease.
Assuntos
Antivirais/uso terapêutico , Medicamentos de Ervas Chinesas/uso terapêutico , Hepatite B/tratamento farmacológico , Hepatite Viral Animal/tratamento farmacológico , Animais , Antivirais/farmacologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA Viral , Patos , Hepatite B/imunologia , Hepatite B/patologia , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B do Pato/efeitos dos fármacos , Vírus da Hepatite B do Pato/genética , Vírus da Hepatite B do Pato/imunologia , Antígenos E da Hepatite B/imunologia , Hepatite Viral Animal/imunologia , Hepatite Viral Animal/patologia , Hepatite Viral Animal/virologia , Humanos , Fígado/efeitos dos fármacos , Fígado/patologia , Medicina Tradicional ChinesaRESUMO
The anti-HBV effect of methyl helicterate (MH), a triterpenoid isolated from the Chinese herb Helicteres angustifolia, was explored both in vitro and in vivo. In the HBV-transfected cell line HepG2.2.15, the secretion of HBsAg/HBeAg, the levels of HBV DNA and cccDNA, and the amount of viral RNA were significantly decreased after treatment with MH for 144h. In addition, MH had no inhibitory effect on the mitochondrial DNA content. In DHBV-infected ducklings, MH significantly reduced the serum DHBV DNA, liver total viral DNA, and cccDNA levels. Furthermore, analysis of the liver pathological changes confirmed the hepatoprotective effect of MH. These results indicate that MH efficiently inhibits HBV replication both in vitro and in vivo and that MH may be a major bioactive ingredient in H. angustifolia.
Assuntos
Antivirais/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Hepatite Viral Animal/tratamento farmacológico , Malvaceae/química , Extratos Vegetais/farmacologia , Triterpenos/farmacologia , Animais , Antivirais/isolamento & purificação , Linhagem Celular , DNA Viral/genética , DNA Viral/isolamento & purificação , Patos , Infecções por Hepadnaviridae/tratamento farmacológico , Vírus da Hepatite B do Pato/genética , Vírus da Hepatite B do Pato/isolamento & purificação , Hepatócitos/virologia , Histocitoquímica , Humanos , Fígado/patologia , Fígado/virologia , Extratos Vegetais/isolamento & purificação , Doenças das Aves Domésticas/tratamento farmacológico , Soro/virologia , Triterpenos/isolamento & purificação , Carga ViralRESUMO
AIMS: To evaluate the anti-hepatitis B virus (anti-HBV) effects and mechanisms of recombinant human serum albumin-interferon-alpha-2b fusion protein (rHSA-IFNalpha-2b) in vitro and in vivo. METHODS: The inhibiting effects on HBV replication were examined in the HepG2 2.2.15 cell line and in ducks, and the expressions of signal transducers and transactivator 1 (STAT1), IFN-stimulated gene factor 3 (ISGF3) and 2',5'-oligoadenylate synthetase 1 (OAS1) were investigated by the reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis. RESULTS: In vitro,at concentrations from 0.075 to 1.2 nmol/l, rHSA-IFNalpha-2b inhibited the releases of extracellular hepatitis B surface antigen, hepatitis B e antigen and HBV DNA in a dose-dependent manner; rHSA- IFNalpha-2b also increased the levels of STAT1, ISGF3 and OAS1. In vivo, rHSA-IFNalpha-2b reduced the levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin and duck hepatitis B virus (DHBV) DNA in the sera of DHBV-infected ducks. CONCLUSIONS: We provide the first evidence that rHSA-IFNalpha-2b significantly inhibits HBV replication in HepG2 2.2.15 cells and in ducks, and that the antiviral effect of rHSA-IFNalpha-2b in vivo is more potent than that of IFNalpha-2b. The anti-HBV mechanism probably operates by triggering the JAK-STAT signaling pathway and increasing the expression of OAS1.
Assuntos
Antivirais/farmacologia , Vírus da Hepatite B do Pato/efeitos dos fármacos , Vírus da Hepatite B do Pato/metabolismo , Vírus da Hepatite B/efeitos dos fármacos , Vírus da Hepatite B/metabolismo , Interferon-alfa/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , 2',5'-Oligoadenilato Sintetase/metabolismo , Alanina Transaminase/sangue , Animais , Aspartato Aminotransferases , Bilirrubina/sangue , Linhagem Celular , DNA/sangue , DNA Viral , Avaliação Pré-Clínica de Medicamentos , Patos , Vírus da Hepatite B do Pato/genética , Humanos , Interferon alfa-2 , Fator Gênico 3 Estimulado por Interferon/metabolismo , Fígado/metabolismo , Fígado/patologia , Distribuição Aleatória , Proteínas Recombinantes , Fator de Transcrição STAT1/metabolismo , Albumina Sérica , Albumina Sérica Humana , Regulação para Cima/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
Total ethanol extract and saponins from Chinese herb radix Astragali (huangqi) have been previously shown to possess anti-hepatitis B virus (HBV) activities in vitro. To identify the active ingredients, we isolated a triterpenoid saponin that was determined to be astragaloside IV. In the human HBV-transfected liver cell line HepG(2) 2.2.15, astragaloside IV effectively suppressed secretion of HBV antigens with inhibition rates of 23.6% for the secretion of Hepatitis B surface antigen (HBsAg) and 22.9% for that of Hepatitis B e antigen (HBeAg) at 100 microg/ml after 9 d of treatment. The inhibitory activity of astragaloside IV on secretion of HBV antigens is more potent than that of 3TC without significant cytotoxicity. In duck hepatitis B virus (DHBV)-infected ducklings, astragaloside IV caused 64.0% inhibition at 120 mg/kg, 49.6% inhibition at 40 mg/kg, and 41.7% inhibition at 10 mg/kg to serum DHBVs after 10 d of treatment, and also reduced serum DHBV DNA levels. Together, our results demonstrate that astragaloside IV possesses potent anti-HBV activity.
Assuntos
Astrágalo/química , Medicamentos de Ervas Chinesas/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Saponinas/farmacologia , Triterpenos/farmacologia , Animais , Carcinoma , Linhagem Celular Tumoral , Relação Dose-Resposta a Droga , Patos , Antígenos de Superfície da Hepatite B/metabolismo , Vírus da Hepatite B do Pato/efeitos dos fármacos , Vírus da Hepatite B do Pato/genética , Antígenos E da Hepatite B/metabolismo , Humanos , Transfecção/métodosRESUMO
AIM: To study the antiviral effect of Oenanthe javanica flavones (OjF) on human hepatoma HepG2.2.15 culture system and duck hepatitis B virus (DHBV) infection. METHODS: (1) After incubation for 24 h, the 2.2.15 cells were treated with different concentrations of OjF for 12 d. The cell alteration was observed by microscope. The presence of HBsAg and HBeAg were measured using the enzyme immunoassay kit after 2.2.15 cells were treated with OjF for 9 d. (2) Ducklings infected with DHBV intravenously were divided into 5 groups and treated with OjF, acyclovir (ACV), and normal saline respectively for 10 d. All the ducklings were bled before, during, and after treatments at different times, and serum levels of DHBV-DNA were detected by a dot-blot hybridization assay. RESULTS: (1) The 50% toxic concentration (TC50) of OjF was 2.28 g/L. The maximum nontoxic concentration (TC0) was 1.00 g/L. In nontoxic concentrations, OjF significantly inhibited HBsAg and HBeAg in 2.2.15 cells after 9 d of treatment (P<0.05, P<0.01). (2) The DHBV-DNA levels decreased significantly after the treatment with 0.50 and 1.00 g/kg of OjF (P<0.01). The inhibition of the peak of viremia was maximum at a dose of 1.00 g/kg and reached 54.3% on d 5 and 64.5% on d 10, respectively. CONCLUSION: The results demonstrate that OjF is a strong inhibitor of HBsAg and HBeAg secretion in 2.2.15 cells and DHBV-DNA levels in the HBV-infected duck model.
Assuntos
Antivirais/farmacologia , DNA Viral/biossíntese , Flavonas/farmacologia , Vírus da Hepatite B do Pato/efeitos dos fármacos , Oenanthe , Animais , Antivirais/isolamento & purificação , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/patologia , Linhagem Celular Tumoral , Replicação do DNA/efeitos dos fármacos , Medicamentos de Ervas Chinesas/isolamento & purificação , Medicamentos de Ervas Chinesas/farmacologia , Flavonas/isolamento & purificação , Infecções por Hepadnaviridae/virologia , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/biossíntese , Vírus da Hepatite B do Pato/genética , Antígenos E da Hepatite B/biossíntese , Hepatite Viral Animal/virologia , Humanos , Fígado/imunologia , Neoplasias Hepáticas/imunologia , Neoplasias Hepáticas/patologia , Oenanthe/química , Plantas Medicinais/químicaRESUMO
OBJECTIVE: To study the antiviral effect of combination therapy with the nucleoside analog lamivudine and famciclovir on duck hepatitis virus (DHBV) in vivo. METHODS: The Chongqing duck hepatitis B model was treated with lamivudine and famciclovir by intragastric administration for 4 weeks. DHBV DNA and DHBsAg in serum were observed by serum dot-blot hybridization and ELISA. ALT and AST in serum were also detected. Histological observation on the duck liver was done simultaneously. The trial was contrasted with a single acyclovir (ACV), famciclovir (FCV), or Lamivudine (3TC). RESULTS: Combination therapy with Lamivudine and famciclovir could significantly reduce the serum DHBV DNA level. After stopping the treatment for 1 week, serum DHBV DNA level did not return significantly. The change of DHBsAg was similar to DHBV DNA. The level of ALT, AST, and the features of liver histopathology in combination-treated ducks were not different from those in control ducks. CONCLUSIONS: The study confirms that combination therapy is superior to single antiviral agent in vivo for ducks with chronic DHBV carrier.
Assuntos
2-Aminopurina/análogos & derivados , 2-Aminopurina/uso terapêutico , Antivirais/uso terapêutico , Infecções por Hepadnaviridae/tratamento farmacológico , Vírus da Hepatite B do Pato , Hepatite Viral Animal/tratamento farmacológico , Lamivudina/uso terapêutico , Animais , DNA Viral/sangue , Modelos Animais de Doenças , Quimioterapia Combinada , Patos , Famciclovir , Vírus da Hepatite B do Pato/efeitos dos fármacos , Vírus da Hepatite B do Pato/genética , Vírus da Hepatite B do Pato/fisiologia , Testes de Sensibilidade MicrobianaRESUMO
A group of myristic acid analogs, designed as alternative substrates for N-myristoyltransferase (NMT), were evaluated against human immunodeficiency virus (HIV), hepatitis B virus (HBV) and duck hepatitis B virus (DHBV) in vitro. Antiviral potency was increased when S or O was substituted for -CH2- in myristic acid and selectivity was affected by the presence and position of the heteroatoms and phenyl groups. A correlation was established among anti-HIV activity, Log P and Log D7.4 and between anti-HIV activity and carbonyl-heteroatom interatomic distances in the myristoyl analogs. 12-Thioethyldodecanoic acid 6 was moderately active (EC50 = 9.37 microM) against HIV-infected T4-lymphocytes (CEM-SS cell line), and it exhibited in vitro activity (EC50 = 17.8 microM) against HBV-producing 2.2.15 cell cultures derived from a human hepatoblastoma cell line (Hep G2). 12-Methoxydodecanoic acid 1 exhibited in vitro activity (EC50 = 20-30 microM) against hepatitis B in the HBV DNA-transfected 2.2.15 cell line. At a concentration of 10 microg/ml, none of the fatty acids significantly inhibited the replication of DHBV in infected hepatocytes.
Assuntos
Fármacos Anti-HIV/farmacologia , Antivirais/farmacologia , HIV-1/efeitos dos fármacos , Vírus da Hepatite B do Pato/efeitos dos fármacos , Vírus da Hepatite B/efeitos dos fármacos , Ácidos Mirísticos/farmacologia , Sequência de Aminoácidos , Animais , Fármacos Anti-HIV/química , Antivirais/química , Linhagem Celular , Fenômenos Químicos , Físico-Química , Avaliação Pré-Clínica de Medicamentos , Ácidos Graxos/química , Ácidos Graxos/farmacologia , HIV-1/genética , HIV-1/fisiologia , Vírus da Hepatite B do Pato/genética , Vírus da Hepatite B do Pato/fisiologia , Vírus da Hepatite B/genética , Vírus da Hepatite B/fisiologia , Humanos , Técnicas In Vitro , Ácidos Mirísticos/química , Relação Estrutura-Atividade , Sulfetos/química , Sulfetos/farmacologia , Proteínas Virais/genética , Proteínas Virais/metabolismo , Replicação Viral/efeitos dos fármacosRESUMO
OBJECTIVE: To assess the efficacy of Yanggan Aoping Mixture (YGAPM) in treating hepatitis B. METHODS: Patients suffered from chronic hepatitis B were treated with YGAPM. Observe their short- and long-term efficacy and the change of serum hepatitis B virus marker. In experiment, the effect of YGAPM in treating rat's liver injury as well as HBV-infected tree shrew and duck HBV-infected ducks was observed. RESULTS: In 79 cases of chronic persistant hepatitis, the markedly effective rate was 60.76%, and follow-up studies on 40 cases, the further rasied to 70.00%. In 73 cases of chronic active hepatitis, the markedly effective rate was 60.27%, and further raised to 62.50% in 32 follow-up cases. In the treatment group, 85 (71.43%) of the 119 cases with HBeAg-positive turned to negative. Whereas in the control group, only 40 (44.94%) of 89 HBeAg-positive cases turned to negative, P < 0.01. Results of experimental study showed that negative conversion rate of tree shrew infected with HBV marker was raised, while infected duck blood with duck HBV DNA was inhibited. Those compared with the control group separately, the difference was remarkably significant. CONCLUSIONS: YGAPM is an effective drug in treating chronic hepatitis B, and it could effectively negative convert the HBV marker.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Hepatite B Crônica/tratamento farmacológico , Adolescente , Adulto , Animais , DNA Viral/sangue , Feminino , Seguimentos , Infecções por Hepadnaviridae/tratamento farmacológico , Vírus da Hepatite B do Pato/genética , Antígenos E da Hepatite B/sangue , Hepatite B Crônica/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Ratos , Ratos Sprague-Dawley , TupaiidaeRESUMO
Eighty-seven cases of chronic persistent hepatitis (CPH) and 29 cases of chronic active hepatitis (CAH) were treated with Astragali Composita (AC), the clinical markedly effective rates were 69.0% and 55.2%, the total effective rates were 90.8% and 89.7% respectively; the sero-negative conversion rates of HBeAg and HBV-DNA were 27.7% and 28.0% respectively, they were higher than that of control group (P < 0.05-0.01). Eight ducks infected hepatitis B virus were treated with AC, sero-negative conversion of DHBV-DNA was seen in 3 ducks, DHBV-DNA levels of duck sera were lower after treatment than that before treatment (P < 0.05); mild inflammatory changes in liver tissues were seen in 3 ducks, its pathologic changes were milder than that of control groups; DHBV-DNA in liver tissues of 2 ducks was negative in situhybridization. The results indicated that AC had obvious effects of protecting liver tissues and some effects of inhibiting sera HBV and HBV replication.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Hepatite B/tratamento farmacológico , Adolescente , Adulto , Animais , DNA Viral/metabolismo , Patos , Feminino , Vírus da Hepatite B do Pato/genética , Antígenos E da Hepatite B/metabolismo , Hepatite Crônica/tratamento farmacológico , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The duck hepatitis B virus model was treated with phyllanthus urinaria of different area and combined with Sophora flavesceus as well as ciprofloxacin once a day for one month, the results indicated: Guangxi and Yunnan Phyllanthus could lower the serum DHBV DNA significantly (P < 0.05), but Chongqing Phyllanthus couldn't. And the amount of serum DHBV DNA rose a week after stopping of Yunnan Phyllanthus. The antiviral effect of Guangxi Phyllanthus combined with ciprofloxacin seems to be strengthened (P < 0.05).
Assuntos
Antivirais/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Infecções por Hepadnaviridae/tratamento farmacológico , Vírus da Hepatite B do Pato/efeitos dos fármacos , Doenças das Aves Domésticas/tratamento farmacológico , Animais , Ciprofloxacina/farmacologia , DNA Viral/sangue , Sinergismo Farmacológico , Patos , Vírus da Hepatite B do Pato/genética , Especificidade da EspécieRESUMO
Extracts of the two traditional Indian herbs, Phyllanthus amarus (P. amarus) and Phyllanthus maderaspatensis (P. maderaspatensis), described by others as useful in the treatment of chronic hepatitis B virus infection were studied for antiviral properties on duck hepatitis B virus infection. One hundred and fourteen ducks infected posthatch with the duck hepatitis B virus (DHBV) were divided into groups at three months of age and treated intraperitoneally with the aqueous, butanol, and alcoholic extracts of these two plants at doses of 25, 50, or 200 mg/kg body weight. Saline-treated animals served as controls. In the ducks negative for DHBV in serum after treatment, we observed replicative intermediates in the liver. There was no definite antiviral property observed in the treated ducks.
Assuntos
Infecções por Hepadnaviridae/tratamento farmacológico , Vírus da Hepatite B do Pato/efeitos dos fármacos , Extratos Vegetais/uso terapêutico , Plantas Medicinais , Animais , Animais Recém-Nascidos , Antivirais/farmacologia , Southern Blotting , Doença Crônica , DNA Viral/sangue , Patos , Infecções por Hepadnaviridae/microbiologia , Vírus da Hepatite B do Pato/genética , Índia , Fígado/microbiologia , Fígado/patologia , Extratos Vegetais/farmacologiaRESUMO
The therapeutic potential of plant extracts of Phyllanthus amarus and Phyllanthus maderas patensis for postexposure prophylaxis against infection by Hepadnaviruses was studied in ducklings infected by the duck hepatitis B virus (DHBV). Forty-four Pekin ducklings were inoculated intraperitoneally with DHBV at 24 hr post-hatch. They were treated by intraperitoneal injection of Phyllanthus amarus (aqueous extract) (100 mg/kg body weight) or Phyllanthus mad eraspatensis (alcoholic extract) (100 mg/kg body weight) for a period of 4 weeks. Infected ducklings treated with saline served as controls. Weekly serum samples obtained before, during, and after treatment were analysed for the presence of DHBV DNA in serum by dot blot hybridisation using alpha 32P-labelled probes. Liver tissue was collected after killing the ducks at various time intervals and was studied for replicative status of the viral DNA and liver histopathology; 17 of 21 ducks were viraemic on completion of treatment with Phyllanthus amarus. At 16 week posttreatment follow-up four of seven animals remained viraemic. Similar results were obtained with Phyllanthus maderaspatensis. There was no alteration in DHBV replication in the liver. No toxicity was observed with this treatment. These observations suggest that Phyllanthus amarus and Phyllanthus maderaspatensis are not useful as therapeutic agents for postexposure prophylaxis against DHBV infection.