Fourteen new geranyl phenyl ethers (1-14) along with three known compounds (15-17) were isolated from Illicium micranthum, and their structures were elucidated by comprehensive spectroscopic methods. Illimicranins A-H (1-8) were characterized as geranyl vanillin ethers, while 9 and 10 were dimethyl acetal derivatives. Illimicranins I and J (11 and 12) were rare geranyl isoeugenol ethers. Illimicranins K and L (13 and 14) represented the first example of geranyl guaiacylacetone ether and geranyl zingerone ether, respectively. Compounds 1, 2 and 15 exhibited anti-HBV (hepatitis B virus) activity against HBsAg (hepatitis B surface antigen) and HBeAg (hepatitis B e antigen) secretion, and HBV DNA replication.
Antiviral Agents/pharmacology* ; Hepatitis B Surface Antigens ; Hepatitis B e Antigens ; Illicium/chemistry* ; Phenyl Ethers

OBJECTIVE: To investigate the effect of extracts with water and alcohol from Radix Trichosanthis on the cell survival and the expression of hepatitis B surface antigen (HBsAg) and hepatitis B e-antigen (HBeAg) in HepG2.2.15 cell supernatant. METHODS: The cell survival rate of HepG2.2.15 cells was detected by MTT assay. The HBsAg and HBeAg in HepG 2.2.15 cell supernatant were evaluated by enzyme linked immunosorbent assay. RESULTS: The water and alcohol soluble extracts from Radix Trichosanthis significantly inhibited the levels of HBsAg and HBeAg in HepG2.2.15 cells in a time-and-concentration-dependent manner. However, the therapeutic index of extracts with water from Radix Trichosanthis was better than that in the alcohol group. CONCLUSION The activity of water-soluble extract from Radix Trichosanthis is stronger on anti-hepatitis B virus than that of the alcohol-soluble extract.
Antiviral Agents ; pharmacology ; Drugs, Chinese Herbal ; classification ; pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens ; biosynthesis ; Hepatitis B e Antigens ; biosynthesis ; Hepatitis B virus ; drug effects ; Humans

In this study, the anti-HBV effects of tea polyphenols (TP) were examined. After cells were exposed to TP for 3, 6, 9 days, amounts of HBsAg, HBeAg and HBV-DNA released into the supernatant of the cultured HepG2 2.2.15 cells were detected. TP, to some extent, inhibited the secretion of HBsAg and strongly suppressed the secretion of HBeAg in a dose-dependent (P<0.01) and time-dependent manner, with 50% maximal inhibitory concentration (IC50) value being 7.34 microg/mL on the 9th day, but the time-dependence was not significant (P=0.051). Expression of HBV-DNA in the supernatant of the cell culture also was significantly decreased in a dose-dependent fashion (P<0.01). The IC50 of TP in inhibiting HBV DNA was 2.54 microg/mL. It concluded that TP possessed potential anti-HBV effects and may be used as a treatment alternative for HBV infection.
Antiviral Agents/*pharmacology ; DNA, Viral/analysis ; Dose-Response Relationship, Drug ; Flavonoids/*pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens/analysis ; Hepatitis B e Antigens/analysis ; Hepatitis B virus/*drug effects ; Inhibitory Concentration 50 ; Phenols/*pharmacology ; Tea/*chemistry

<b>OBJECTIVEb>To evaluate the biological activity of polysaccharide of Cipangopaludina chinensis (PCC) against HBV in vitro.

<b>METHODb>HepG2 2. 2. 15 cells were taken as the in vitro experimental model. The cell toxicity was observed by MTT. PCC of different safe concentrations and positive control medicine 3TC were added into the cells. Cell control without medicine was set at the same time. Cultural supernatants were collected at 9 d. HBsAg and HBeAg in cultural supernatants were tested by ELISA. The content of HBV-DNA was detected by TaqMan probe fluorescence quantitative PCR.

<b>RESULTb>TC0 and TC50 of PCC in HepG2 2. 2. 15 cell culture were 1 g . L-1 and >10 g . L-1, respectively, suggesting low toxicity in cells. IC50 of PCC in HepG2 2. 2. 15 cells HBsAg and HBeAg were 0. 501, 0. 401 g. L-1, with SI being >19.96 and >24. 94, respectively. PCC could effectively inhibit the secretion of HBsAg and HBeAg, and have a better effect on HBeAg than on HBsAg. PCC had a significant inhibitory effect on HBV-DNA in HepG2 2. 2. 15 cells at concentrations of 0. 1, 1 g . L-1 P <0.05).

<b>CONCLUSIONb>PCC has the effect against HBV activity in vitro to some extent, with low toxicity, thereby having a good prospect for application.

Animals ; Antiviral Agents ; adverse effects ; pharmacology ; DNA, Viral ; metabolism ; Hep G2 Cells ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; drug effects ; metabolism ; Humans ; Polysaccharides ; adverse effects ; pharmacology ; Snails ; chemistry

<b>OBJECTIVEb>To investigate the effect of synthetic non-methylated CpG-ODN combined with recombined HBsAg on the phenotype, function and the expression level of nucleic transcription factor NF-kappa B (NF-kB) and AP-1 of monocyte-derived dendritic cells (DC) in patients with chronic hepatitis B (CHB).

<b>METHODSb>Purified monocytes were isolated from the peripheral blood of CHB patients and healthy volunteers and cultured with human granulocyte-monocyte colony stimulating factor added together with interleukin-4. On the fifth day of culture, CpG-ODN, HBsAg and other reagents, such as TNF alpha and PBS, were added to the medium, and 18 hours later cells were collected for the detection of surface molecules (HLA-DR/CD86/CD1a). IL-12p70 levels in the supernatant and stimulating capacity to allogenic T lymphocytes were detected. The nucleic proteins of NF-kB and AP-1 in DC were extracted and purified for the gel shift assay.

<b>RESULTSb>Compared with those of the PBS group, the expression rates of HLA-DR of DC treated with CpG-ODN and/or HBsAg were obviously increased. Both the IL-12p70 level and the stimulating capacity of DC to allogenic T lymphocytes in mixed lymphocyte reaction increased significantly in the CpG-ODN group and in the CpG-ODN/HBsAg combination group (P less than 0.01 and 0.05, respectively). The expression rates of CD1a were raised only in the CpG-ODN/HBsAg combination group. Three kinds of immunological adjuvants, TNF alpha, CpG-ODN and CpG-ODN/HBsAg enhanced the expression of nucleic NF-kB and inhibited the expression of AP-1 in DC.

<b>CONCLUSIONb>CpG-ODN, like TNF alpha, has remarkable stimulatory effect on the impaired phenotype and function of monocyte-derived DC in patients with CHB; CpG-ODN and HBsAg have a synergetic effect in increasing the antigen presenting function. The regulating ability of CpG-ODN and TNF alpha on the expression levels of NF-kB and AP-1 might be one of the mechanisms of their immunostimulatory effects on DC of the CHB patients.

Adjuvants, Immunologic ; pharmacology ; Adult ; Cells, Cultured ; Dendritic Cells ; metabolism ; HLA-DR Antigens ; metabolism ; Hepatitis B Surface Antigens ; pharmacology ; Hepatitis B, Chronic ; metabolism ; Humans ; Male ; NF-kappa B ; metabolism ; Oligodeoxyribonucleotides ; pharmacology ; Recombinant Fusion Proteins ; Transcription Factor AP-1 ; metabolism

CD8-Positive T-Lymphocytes ; metabolism ; Case-Control Studies ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Lymphocytes ; immunology ; Oligodeoxyribonucleotides ; pharmacology

<b>OBJECTIVEb>To investigate the ability of human dendritic cells (DCs) inducing specific T lymphocyte response and inhibit the expression of HBeAg and HBsAg in 2.2.15 cell culture supernatant.

<b>METHODSb>DCs were prepared from peripheral blood mononuclear cells induced with granulocyte macrophage colony-stimulating factor(GM-CSF) and interleukin 4. DCs was impulsed with pure HBsAg before DCs maturation and cocultured with self-blood T lymphocyte, while DCs without pure HBsAg stimulated group, T lymphocyte group and only T lymphocyte group were prepared as control group. The culture supernatant of 2.2.15 cell with stimulated T lymphocytes was collected on day 1, day 3, day 5 and day 7, respectively. The expressed levels of HBeAg and HBsAg were detected by ELISA method.

<b>RESULTSb>DCs after antigen stimulation had a strong ability to present antigen and induce immune activation, DCs after loading with antigen in normal control and chronic hepatitis patients group had stronger stimulative ability for T lymphocytes proliferation than that of DCs without loading with antigen and only T lymphocyte group(P less than 0.01). The stimulating ability of DCs had a positive correlation to the dosage of loaded antigen; CTLs produced as a result of DCs stimulation had a specific inhibitive effect on the expression of HBeAg in 2.2.15 cell supernatant,but not on the expression of HbsAg.

<b>CONCLUSIONb>Human dendritic cells stimulated with HBsAg in vitro can efficiently present antigen and stimulate the production of specific CTLs to HBV, which can efficiently inhibit the expression of HBeAg in 2.2.15 cell supernatant- DC vaccine may become an antiviral therapy strategy for chronic hepatitis B virus infected patients in future.

Adolescent ; Adult ; Antigen Presentation ; Cells, Cultured ; Child ; Dendritic Cells ; drug effects ; immunology ; Female ; Hepatitis B Surface Antigens ; pharmacology ; Hepatitis B, Chronic ; immunology ; Humans ; Lymphocyte Activation ; Male ; Middle Aged

<b>OBJECTIVEb>To evaluate the inhibition effects of DNAzymes specific to Hepatitis B Virus(HBV) s gene and e gene on the expressions of Hepatitis B surface antigen(HBsAg) and Hepatitis B e antigen(HBeAg).

<b>METHODSb>DNAzymes DrzBS and DrzBC specific to HBV s gene ORF A157UG and e gene ORF A1816UG, respectively, were designed and synthesized. The inhibition effects of DrzBS or DrzBC on the expressions of HBV s and e genes were observed in 2.2.15 cells.

<b>RESULTSb>The expression of HBV s or e genes was dramatically depressed after 2.2.15 cells treated by DrzBS or DrzBC. The concentration for effective inhibition was within 0.1-2.5 micromol/L and the inhibition showed a dose dependence within that concentration range. The maximum inhibition was 94.2% and 91.8% for DrzBS and DrzBC, respectively. The inhibition was maintained for 72 hours. The efficiency of inhibiting HbsAg, HbeAg in 2.2.15 cells by DrzBS, DrzBC was higher than that by antisense oligonucleotides for the same target genes. The concentrations for effective inhibition of the DNAzymes were at least 10-fold lower compared with antisense oligonucleotides. Neither inhibition on the replication of HBV DNA nor toxicity to 2.2.15 cells was observed.

<b>CONCLUSIONb>DrzBS and DrzBC can highly block the expressions of HBV s gene and e gene in 2.2.15 HBV cell model and are proved a specific and effective anti-HBV gene therapeutic means.

DNA, Catalytic ; pharmacology ; therapeutic use ; DNA, Viral ; analysis ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Genetic Therapy ; Hepatitis B ; therapy ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B e Antigens ; genetics

<b>OBJECTIVEb>To explore the activities of Composite Artemisia Capillaris Tablet (CACT) against hepatitis B virus replication in vitro.

<b>METHODSb>By means of radioimmunoassay (RIA), Dot blot and Southern blot, the surface and e antigen production of 2.2.15 cells, HBV DNA in 2.2.15 cell culture medium and that in 2.2.15 cells were examined respectively.

<b>RESULTSb>HBsAg, HBeAg values of 2.2.15 cells treated by CACT were lower than those of the control, the HBV DNA quantities in culture medium and in 2.2.15 cells decreased as compared with those cells with no treatment by CACT given to them.

<b>CONCLUSIONb>CACT could inhibit HBV DNA replication, showing its potential antiviral activity in hepatitis B treatment.

Cell Line, Tumor ; DNA, Viral ; antagonists & inhibitors ; Hepatitis B Surface Antigens ; metabolism ; Hepatitis B e Antigens ; metabolism ; Hepatitis B virus ; genetics ; immunology ; physiology ; Humans ; Medicine, Chinese Traditional ; Plant Preparations ; administration & dosage ; pharmacology ; toxicity ; Radioimmunoassay ; Tablets ; Virus Replication ; drug effects

To verify the effect of echinacoside on replication and antigen expression of hepatitis B virus (HBV) by using HBV-transfected HepG2. 2. 15 cells as the in vitro model. The ELISA method was used to determine HBeAg and HBsAg levels in cellular supernatants. The effect of echinacoside on HBV replication was studied by using HBV transgenic mice as the in vivo model. First of all, the HBV DNA level in hepatic tissues was quantified with PCR method. Meanwhile, the serum transaminase levels and hepatic pathological changes were also evaluated. Subsequently, HBV transgenic mice were divided into five groups: the control group, the lamivudine group (50 mg · kg(-1)) and echinacoside high, medium and low dose group (50, 25 and 12.5 mg · kg(-1)). The mice were orally administered with drugs once per day for 30 days. At the 31st day, the mice serum was separated to measure HBsAg, HBeAg and HBV DNA. Additionally, the liver HBV DNA level and histopathological change were detected. The results indicated that echinacoside at 50 and 100 mg · L(-1) suppressed significantly HBsAg and HBeAg expressions on the sixth day, with the maximum inhibition ratios of 42.68% and 46.29%; And echinacoside at 100 mg · L(-1) also showed an inhibitory effect on HBV DNA. Besides, echinacoside at 50 mg · kg(-1) inhibited significantly HBsAg and HBeAg expressions of HBV transgenic mice, with the inhibition ratios of 42.82% and 29.12%, and reduced markedly the serum HBV DNA level in HBV transgenic mice. In conclusion, the study suggested that echinacoside has a strong effect against HBV replication and antigen expression.
Animals ; DNA, Viral ; blood ; Female ; Glycosides ; pharmacology ; Hep G2 Cells ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Virus Replication ; drug effects