AIM： To investigate the antiviral effect of beta-L- enantiomer of 2;3＇-didehydro-2＇,3″-dideoxyadenosine （13-L-D4A） on 2.2.15 cells transfected with the hepatitis B virus （HBV） genome.METHODS： Lamivudine （3TC...AIM： To investigate the antiviral effect of beta-L- enantiomer of 2;3＇-didehydro-2＇,3″-dideoxyadenosine （13-L-D4A） on 2.2.15 cells transfected with the hepatitis B virus （HBV） genome.METHODS： Lamivudine （3TC） as a positive control. Then, HBV DNA in treated 2.2.15 cells and the Hepatitis B surface antigen （HBsAg） in the culture supernatants were detected to determine the inhibitory effect of β-L- D4A. At the same time, 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） was used to detect the survival ratio of 2.2.15 cells.RESULTS： β-L-D4A has a dose-dependent inhibitory effect on HBV DNA replication; this effect was apparent when the concentration was above 1 mol/L. When β-L- D4A was at the highest concentration, 100 mol/L, the HBsAg inhibition ratio was above 50%. The Therapeutic index （TI） of β-L-D4A was above 2.1.CONCLUSION： β-L-D4A has a dose-dependent inhibitory effect on the replication of HBV DNA and the secretion of HBsAg at low toxicity,展开更多
This study aimed to determine the effects of black tea polyphenols on gene expression in hepatocellular cancer cells.The total RNA from HepG_(2) hepatocellular cancer cells treated with black tea polyphenols was subje...This study aimed to determine the effects of black tea polyphenols on gene expression in hepatocellular cancer cells.The total RNA from HepG_(2) hepatocellular cancer cells treated with black tea polyphenols was subjected to Human 14K cDNA microarray analysis.Real-time PCR and Western blot analysis were conducted to verify microarray data.Black tea polyphenols treatment at the dose of 20 mg/L,40 mg/L or 80 mg/L for one to three days inhibited the growth of HepG_(2) cells in a dose and time dependent manner.A total of 48 genes showed more than two-fold change after black tea polyphenols treatment,including 17 upregulated genes and 31 downregulated genes,and they were involved in the regulation of cell growth,cell cycle,apoptosis,signaling,angiogenesis,tumor invasion and metastasis.Real-time PCR analysis of the selected genes showed that their mRNA expression changes were consistent with the microarray data.In addition,Western blot analysis of the selected genes showed that their protein expression changes were consistent with mRNA expression.In conclusion,gene expression profiles provide comprehensive molecular mechanisms by which black tea polyphenols exerts growth inhibition effects on cancer cells.The novel molecular targets identified in this study may be further exploited as therapeutic strategies for hepatocellular cancer.展开更多
Objective:To investigate capacity of Hyptis suaveolens(H.suaveolens) methanol extract as an antioxidant to protect against carbon tetrachloride(CCl_4)-indueed oxidative stress,hepatotoxicitv in Albino Wistar rats and ...Objective:To investigate capacity of Hyptis suaveolens(H.suaveolens) methanol extract as an antioxidant to protect against carbon tetrachloride(CCl_4)-indueed oxidative stress,hepatotoxicitv in Albino Wistar rats and cytoprotective effect of hydrogen peroxide(H_2O_2) induced cell death in HepG_2 cell line.Methods:Two different doses of methanol extract of H.suaveolens were evaluated for the hepatoprotective activity against carbon tetrachloride(CCl_4) induced hepatotoxicitv in rats.Animals in GroupⅠ:served as control,groupⅡ:H.suaveolens(100 mL/ kg b.w),groupⅢ:H.suaveolens(50 mL/kg b.w) + CCl_4(1 mg/kg),groupⅣ:H.suaveolens(100 mL/kg b.w) + CCl_4(1 ml/kg)and groupⅤ:CCl_4(1 mL/kg).Histopathologic changes of liver were also evaluated.Cytotoxicity was also determined by 3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide(MTT) assay.Results:Oral sigle dose treatment of CCl_4 produced a marked elevation in the serum levels of aspartate transaminase(AST),alanine transaminase(ALT), alkaline phosphatase(ALP) and Lactate dehydrogenase(LDH).Histopathological analysis of the liver of CCl_4-induced rats revealed marked liver cell necrosis with inflammatory collections that were conformed to increase in the levels of SOD,GSH,GST,GR and LPO.Treatment with H_2O_2 significantly induced death of HepG_2 cell.Pretreatment with H.suaveolens methanol extract inhibited or attenuated H_2O_2 induced cytotoxicity.Conclusions:This study shows that H.suaveolens methanol extract can be proposed to protect the liver against CCl_4-induced oxidative damage in rats and protect the cells against H_2O_2-induced oxidative damage in HepG_2 cells.The hepatoprotective and cytoprotective effects might be correlated with its antioxidant and free radical scavenger effects.展开更多
To investigate the effects of the polysaccharides from Spirulina platensis (PSP) on hepatitis B virus (HBV), the cytotoxic effect of PSP was assessed by choosing the maximal concentration of PSP without cytotoxic effe...To investigate the effects of the polysaccharides from Spirulina platensis (PSP) on hepatitis B virus (HBV), the cytotoxic effect of PSP was assessed by choosing the maximal concentration of PSP without cytotoxic effect on HepG2 2.2.15 cell line for further experiments. Four concentrations (400-50 μg/ml) of PSP were adopted to treat the cells, and the supernatants or cells were collected after 24, 48, 72 and 144 h respectively. HBsAg and HBeAg in the supernatants of cell cultures were tested with ELISA and copies of HBV DNA in supernatants were measured by real-time fluorescence quantitative PCR (FQ-PCR). Meanwhile, DNA/RNA hybridization was performed to evaluate the expression of IFN-α receptor (IFN-αR) on the cells. The experimental results showed that the secretion of HBV antigens decreased under the influence of PSP in a dose and time-dependent manner. PSP in concentration of 400 μg/ml could significantly decrease the secretion of HBsAg in 24 h. Although no obvious effect was observed on the secretion of HBeAg at that time, the inhibitory effects were observed in a dose-dependent manner from 48 to 144 h. In addition, the copies of HBV DNA were declined under the influence of PSP in the same manner, moreover, the maximal suppressive effect of PSP in concentration of 400 μg/ml was as great as that of lamivudine. The expression of IFN-αR was much higher in PSP-treated cells than that of the un-treated cells also in dose and time-dependent manner. It is concluded that PSP in non-cytotoxic concentration not only significantly decreases the secretion of the HBV antigens and the replication of HBV DNA, but also increases the expression of IFN-αR on HepG2 2.2.15 cell line. The results of the present investigation strongly support the notion that PSP exerts the anti-HBV effect both through enhancing the anti-HBV immunity and acting on HBV directly.展开更多
Background and Aims:Hepatitis B virus(HBV)infection is a major risk factor for cirrhosis and liver cancer,and its treatment continues to be difficult.We previously demonstrated that a dopamine analog inhibited the pac...Background and Aims:Hepatitis B virus(HBV)infection is a major risk factor for cirrhosis and liver cancer,and its treatment continues to be difficult.We previously demonstrated that a dopamine analog inhibited the packaging of pregenomic RNA into capsids.The present study aimed to determine the effect of dopamine on the expressions of hepatitis B virus surface and e antigens(HBsAg and HBeAg,respectively)and to elucidate the underlying mechanism.Methods:We used dopamine-treated HBVinfected HepG2.2.15 and NTCP-G2 cells to monitor HBsAg and HBeAg expression levels.We analyzed interferon-stimulated gene 15(ISG15)expression in dopamine-treated cells.We knocked down ISG15 and then monitored HBsAg and HBeAg expression levels.We analyzed the expression of Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway factors in dopamine-treated cells.We used dopamine hydrochloride-treated adeno-associated virus/HBV-infected mouse model to evaluate HBV DNA,HBsAg,and HBeAg expression.HBV virus was collected from HepAD38.7 cell culture medium.Results:Dopamine inhibited HBsAg and HBeAg expression and upregulated ISG15 expression in HepG2.2.15 and HepG2-NTCP cell lines.ISG15 knockdown increased HBsAg and HBeAg expression in HepG2.2.15 cells.Dopamine-treated cells activated the JAK/STAT pathway,which upregulated ISG15 expression.In the adeno-associated virus-HBV murine infection model,dopamine downregulated HBsAg and HBeAg expression and activated the JAK-STAT/ISG15 axis.Conclusions:Dopamine inhibits the expression of HBsAg and HBeAg by activating the JAK/STAT pathway and upregulating ISG15 expression.展开更多
Objective To evaluate the effect of 8-epi-kingiside (8-Epik) derived from the buds of Jasminum officinale var. grandiflorum (JOG) on hepatitis B virus (HBV) replication in HepG2 2.2.15 cell line in vitro and duck hepa...Objective To evaluate the effect of 8-epi-kingiside (8-Epik) derived from the buds of Jasminum officinale var. grandiflorum (JOG) on hepatitis B virus (HBV) replication in HepG2 2.2.15 cell line in vitro and duck hepatitis B virus (DHBV) replication in ducklings in vivo. Methods The concentration of extracellular hepatitis B e antigen and hepatitis B surface antigen (HBsAg) in cell culture medium was determined by ELISA, respectively. The anti-HBV effects of 8-Epik were also demonstrated in the model of DHBV. 8-Epik was ip given (20, 40, and 80 mg/kg, twice daily) to the DHBV-infected ducklings for 10 d. The isotonic saline liquid diet was ip given as negative control and Lamivudine (50 mg/kg, twice daily) was given as positive control. DHBV DNA was measured at days 0 (T0), 5 (T5), 10 (T10), and day 3 after cessation of treatment (P3) by dot blotting. Results 8-Epik effectively blocked HBsAg secretion in HepG2 2.2.15 cells in a dose-dependent manner [IC 50 = (19.4 ± 1.04) μg/mL]. 8-Epik (40 or 80 mg/kg, ip, twice daily) also reduced viremia in DHBV-infected ducks. Conclusion Therefore, 8-Epik is warranted as a potential therapeutic agent for HBV infection.展开更多
基金Grants from the National Natural Science Foundation of China, Key Program, No. 30330680
文摘AIM： To investigate the antiviral effect of beta-L- enantiomer of 2;3＇-didehydro-2＇,3″-dideoxyadenosine （13-L-D4A） on 2.2.15 cells transfected with the hepatitis B virus （HBV） genome.METHODS： Lamivudine （3TC） as a positive control. Then, HBV DNA in treated 2.2.15 cells and the Hepatitis B surface antigen （HBsAg） in the culture supernatants were detected to determine the inhibitory effect of β-L- D4A. At the same time, 3-（4,5-dimethylthiazol-2-yl）-2,5- diphenyltetrazolium bromide （MTT） was used to detect the survival ratio of 2.2.15 cells.RESULTS： β-L-D4A has a dose-dependent inhibitory effect on HBV DNA replication; this effect was apparent when the concentration was above 1 mol/L. When β-L- D4A was at the highest concentration, 100 mol/L, the HBsAg inhibition ratio was above 50%. The Therapeutic index （TI） of β-L-D4A was above 2.1.CONCLUSION： β-L-D4A has a dose-dependent inhibitory effect on the replication of HBV DNA and the secretion of HBsAg at low toxicity,
基金supported by grants from the Science and Technology Foundation of Health and Family Planning Commission of Hunan Province(No.C2019048)Science and Technology Bureau of Changsha(No.ZD1702024).
文摘This study aimed to determine the effects of black tea polyphenols on gene expression in hepatocellular cancer cells.The total RNA from HepG_(2) hepatocellular cancer cells treated with black tea polyphenols was subjected to Human 14K cDNA microarray analysis.Real-time PCR and Western blot analysis were conducted to verify microarray data.Black tea polyphenols treatment at the dose of 20 mg/L,40 mg/L or 80 mg/L for one to three days inhibited the growth of HepG_(2) cells in a dose and time dependent manner.A total of 48 genes showed more than two-fold change after black tea polyphenols treatment,including 17 upregulated genes and 31 downregulated genes,and they were involved in the regulation of cell growth,cell cycle,apoptosis,signaling,angiogenesis,tumor invasion and metastasis.Real-time PCR analysis of the selected genes showed that their mRNA expression changes were consistent with the microarray data.In addition,Western blot analysis of the selected genes showed that their protein expression changes were consistent with mRNA expression.In conclusion,gene expression profiles provide comprehensive molecular mechanisms by which black tea polyphenols exerts growth inhibition effects on cancer cells.The novel molecular targets identified in this study may be further exploited as therapeutic strategies for hepatocellular cancer.
基金supported by Ministry of Human Resource Developmenl and University Grant Commission Under Institution of Excellence Scheme awarded to the University of Mysore
文摘Objective:To investigate capacity of Hyptis suaveolens(H.suaveolens) methanol extract as an antioxidant to protect against carbon tetrachloride(CCl_4)-indueed oxidative stress,hepatotoxicitv in Albino Wistar rats and cytoprotective effect of hydrogen peroxide(H_2O_2) induced cell death in HepG_2 cell line.Methods:Two different doses of methanol extract of H.suaveolens were evaluated for the hepatoprotective activity against carbon tetrachloride(CCl_4) induced hepatotoxicitv in rats.Animals in GroupⅠ:served as control,groupⅡ:H.suaveolens(100 mL/ kg b.w),groupⅢ:H.suaveolens(50 mL/kg b.w) + CCl_4(1 mg/kg),groupⅣ:H.suaveolens(100 mL/kg b.w) + CCl_4(1 ml/kg)and groupⅤ:CCl_4(1 mL/kg).Histopathologic changes of liver were also evaluated.Cytotoxicity was also determined by 3,(4,5-dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide(MTT) assay.Results:Oral sigle dose treatment of CCl_4 produced a marked elevation in the serum levels of aspartate transaminase(AST),alanine transaminase(ALT), alkaline phosphatase(ALP) and Lactate dehydrogenase(LDH).Histopathological analysis of the liver of CCl_4-induced rats revealed marked liver cell necrosis with inflammatory collections that were conformed to increase in the levels of SOD,GSH,GST,GR and LPO.Treatment with H_2O_2 significantly induced death of HepG_2 cell.Pretreatment with H.suaveolens methanol extract inhibited or attenuated H_2O_2 induced cytotoxicity.Conclusions:This study shows that H.suaveolens methanol extract can be proposed to protect the liver against CCl_4-induced oxidative damage in rats and protect the cells against H_2O_2-induced oxidative damage in HepG_2 cells.The hepatoprotective and cytoprotective effects might be correlated with its antioxidant and free radical scavenger effects.
文摘To investigate the effects of the polysaccharides from Spirulina platensis (PSP) on hepatitis B virus (HBV), the cytotoxic effect of PSP was assessed by choosing the maximal concentration of PSP without cytotoxic effect on HepG2 2.2.15 cell line for further experiments. Four concentrations (400-50 μg/ml) of PSP were adopted to treat the cells, and the supernatants or cells were collected after 24, 48, 72 and 144 h respectively. HBsAg and HBeAg in the supernatants of cell cultures were tested with ELISA and copies of HBV DNA in supernatants were measured by real-time fluorescence quantitative PCR (FQ-PCR). Meanwhile, DNA/RNA hybridization was performed to evaluate the expression of IFN-α receptor (IFN-αR) on the cells. The experimental results showed that the secretion of HBV antigens decreased under the influence of PSP in a dose and time-dependent manner. PSP in concentration of 400 μg/ml could significantly decrease the secretion of HBsAg in 24 h. Although no obvious effect was observed on the secretion of HBeAg at that time, the inhibitory effects were observed in a dose-dependent manner from 48 to 144 h. In addition, the copies of HBV DNA were declined under the influence of PSP in the same manner, moreover, the maximal suppressive effect of PSP in concentration of 400 μg/ml was as great as that of lamivudine. The expression of IFN-αR was much higher in PSP-treated cells than that of the un-treated cells also in dose and time-dependent manner. It is concluded that PSP in non-cytotoxic concentration not only significantly decreases the secretion of the HBV antigens and the replication of HBV DNA, but also increases the expression of IFN-αR on HepG2 2.2.15 cell line. The results of the present investigation strongly support the notion that PSP exerts the anti-HBV effect both through enhancing the anti-HBV immunity and acting on HBV directly.
基金supported by a grant from the National Natural Science Foundation of China(82170612)Guangzhou Science and Technology Program Key Projects(2023B01J1007)National Natural Science Foundation of China(No.81870597).
文摘Background and Aims:Hepatitis B virus(HBV)infection is a major risk factor for cirrhosis and liver cancer,and its treatment continues to be difficult.We previously demonstrated that a dopamine analog inhibited the packaging of pregenomic RNA into capsids.The present study aimed to determine the effect of dopamine on the expressions of hepatitis B virus surface and e antigens(HBsAg and HBeAg,respectively)and to elucidate the underlying mechanism.Methods:We used dopamine-treated HBVinfected HepG2.2.15 and NTCP-G2 cells to monitor HBsAg and HBeAg expression levels.We analyzed interferon-stimulated gene 15(ISG15)expression in dopamine-treated cells.We knocked down ISG15 and then monitored HBsAg and HBeAg expression levels.We analyzed the expression of Janus kinase(JAK)/signal transducer and activator of transcription(STAT)pathway factors in dopamine-treated cells.We used dopamine hydrochloride-treated adeno-associated virus/HBV-infected mouse model to evaluate HBV DNA,HBsAg,and HBeAg expression.HBV virus was collected from HepAD38.7 cell culture medium.Results:Dopamine inhibited HBsAg and HBeAg expression and upregulated ISG15 expression in HepG2.2.15 and HepG2-NTCP cell lines.ISG15 knockdown increased HBsAg and HBeAg expression in HepG2.2.15 cells.Dopamine-treated cells activated the JAK/STAT pathway,which upregulated ISG15 expression.In the adeno-associated virus-HBV murine infection model,dopamine downregulated HBsAg and HBeAg expression and activated the JAK-STAT/ISG15 axis.Conclusions:Dopamine inhibits the expression of HBsAg and HBeAg by activating the JAK/STAT pathway and upregulating ISG15 expression.
基金Nature Science Foundation of Hebei Province of China (C2010001354)
文摘Objective To evaluate the effect of 8-epi-kingiside (8-Epik) derived from the buds of Jasminum officinale var. grandiflorum (JOG) on hepatitis B virus (HBV) replication in HepG2 2.2.15 cell line in vitro and duck hepatitis B virus (DHBV) replication in ducklings in vivo. Methods The concentration of extracellular hepatitis B e antigen and hepatitis B surface antigen (HBsAg) in cell culture medium was determined by ELISA, respectively. The anti-HBV effects of 8-Epik were also demonstrated in the model of DHBV. 8-Epik was ip given (20, 40, and 80 mg/kg, twice daily) to the DHBV-infected ducklings for 10 d. The isotonic saline liquid diet was ip given as negative control and Lamivudine (50 mg/kg, twice daily) was given as positive control. DHBV DNA was measured at days 0 (T0), 5 (T5), 10 (T10), and day 3 after cessation of treatment (P3) by dot blotting. Results 8-Epik effectively blocked HBsAg secretion in HepG2 2.2.15 cells in a dose-dependent manner [IC 50 = (19.4 ± 1.04) μg/mL]. 8-Epik (40 or 80 mg/kg, ip, twice daily) also reduced viremia in DHBV-infected ducks. Conclusion Therefore, 8-Epik is warranted as a potential therapeutic agent for HBV infection.
