[Effect and mechanism of danshensu on hepatitis B virus reverse transcriptase and antigen expression].

MTT assay was used in this study to investigate the inhibitory effect of danshensu on the activity of 2.2.15 cells among human hepatoma cell line (HepG2); indirect fluorescence labeling method was used to measure the changes of reactive oxygen levels in the cells; ELISA method was used to determine hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) levels in cellular supernatants; HBV DNA level was measured with fluorogenic quantitative PCR method. The inhibitory effect of danshensu on HBV RT(hepatitis B virus reverse transcriptase) was studied by using enzyme inhibition dynamics, and the effect of danshensu on secondary structure of HBV reverse transcriptase was monitored by using circular dichroism. The results showed that danshensu had a good inhibitory effect on the growth of HepG2.2.15 cells, with a half inhibitory concentration (IC50) of (15.35±2.43) µmol•L⁻¹; danshensu could significantly inhibit HBsAg and HBeAg expressions, and showed an inhibitory effect on HBV DNA replication. In addition, danshensu was an effective inhibitor for HBV reverse transcriptase [IC50 (21.32±2.43) µmol•L⁻¹]. The fluorescence labeling results showed that the reactive oxygen levels in the cells were increased with the increase of danshensu concentration. Circular dichroism analysis showed that danshensu could induce partial change of conformation of HBV reverse transcriptase and gradually increased α-helical content. These results indicated that danshensu could make the structure of the enzyme become closer by binding to HBV reverse transcriptase, which was not conducive to the formation of the active center, so it could finally decrease the activity of HBV reverse transcriptase. Such decrease in enzyme activity would directly affect the HBV DNA replication, and combined with the decrease of the antigen levels, the effect of danshensu on HBV was increased.

Urinary metabonomics for diagnosis of depression in hepatitis B virus-infected patients.

BACKGROUND: Depression is concomitantly presented in Hepatitis B Virus (HBV)-infected patients and decreases these patients' quality of life. However, there are no laboratory-based methods to objectively diagnose this disorder. OBJECTIVES: The aim of this study was to investigate the alteration of urinary metabolites in depressed HBV-infected patients. PATIENTS AND METHODS: In this study, 81 depressed HBV-infected patients, 68 non-depressed HBV-infected patients and 64 Healthy Controls (HC) were recruited. A nuclear magnetic resonance (NMR)-based urinary metabonomic method was used to characterize the urinary metabolic profiling of depressed and non-depressed subjects. RESULTS: Seventeen differential urinary metabolites responsible for discriminating depressed HBV-infected patients from non-depressed HBV-infected patients and HC were identified. Among these metabolites, pyruvate, isobutyrate, N-methylnicotinamide, α-hydroxybutyrate, acetoacetate and malonate were identified as potential biomarkers for diagnosing depression in HBV-infected patients. A combined panel of these potential biomarkers could effectively discriminate depressed HBV-infected patients from non-depressed HBV-infected patients and HC, with an average accuracy of 89.6% in the training set and a predictive accuracy of 86.4% in the test set. CONCLUSIONS: These findings suggest that NMR-based urinary metabonomics approach might be a useful tool for the clinical diagnosis of depression in HBV-infected patients and the six potential biomarkers could be helpful for developing an objective diagnostic method. Limited by the number of recruited subjects, future studies are required to validate our conclusions.

[Correlation of the expression of NF-κB p65 and hepatic fibrosis in hepatitis patients].

OBJECTIVE: To explore the relationship between the expression of NF-κB p65 and hepatic fibrosis in chronic hepatitis B (CHB) patients. METHODS: Sixty CHB patients with hepatic fibrosis underwent liver biopsy to determine the stages of liver fibrosis (S0-S4). The distribution and expression of collagens I, III and NF-κB p65 in different stages of fibrosis in liver tissue were observed by immunohistochemistry and the results analyzed statistically. RESULTS: The expression of NF-κB p65 was positively correlated with the stage of hepatic fibrosis. That was S4 > S3 > S2 > S1 (S0) (P < 0.01). And it was also positively correlated with the expression of collagens I and III (P < 0.01). CONCLUSION: The elevated expression of NF-κB p65 is closely associated with the occurrence and development of hepatic fibrosis. Its mechanism is probably related with the increased secretion of collagens I and III after the activation of hepatic stellate cell.

[Effects of oxymatrine on serum cytokines and hepatic fibrotic indexes in patients with chronic hepatitis B].

OBJECTIVE: To study the effect of oxymatrine on serum cytokines and hepatic fibrotic indexes in patients with chronic hepatitis B (CHB). METHODS: Eighty-two CHB patients were randomly assigned to the control group treated with Western routine therapy and the treated group treated by Western routine therapy plus oxymatrine. The treatment course was 24 weeks. Another group with 20 healthy subjects was set as the normal control group. The serum levels of transforming growth factor-beta1 (TGF-beta1), interleukin-6 (IL-6), hyaline acid (HA), collagen type IV (C-IV) and laminin (LN) were measured by ELISA and RIA before and after treatment. RESULTS: The serum levels of TGF-beta1, IL-6, HA, C-IV and LN in CHB patients were significantly higher than those in the normal control group (P < 0.01). After 24-week treatment with oxymatrine, these abnormal indexes in the treated group were significantly lowered, as compared with those before treatment and in the control group, the differences were significant (all P < 0.01). CONCLUSION: Oxymatrine could lower the levels of cytokines, including TGF-beta1, IL-6, etc. in patients with CHB, which might be one of the mechanisms of its action in reversing liver fibrosis.

Chronic hepatitis B serum promotes apoptotic damage in human renal tubular cells.

AIM: To investigate the effect of the serum of patients with chronic hepatitis B (CHB) on apoptosis of renal tubular epithelial cells in vitro and to study the role of hepatitis B virus (HBV) and transforming growth factor-beta (1) (TGF-beta (1)) in the pathogenesis of hepatitis B virus associated glomerulonephritis (HBV-GN). METHODS: The levels of serum TGF-beta(1) were measured by specific enzyme linked immunosorbent assay (ELISA) and HBV DNA was tested by polymerase chain reaction (PCR) in 44 patients with CHB ,and 20 healthy persons as the control. The normal human kidney proximal tubular cell (HK-2) was cultured together with the sera of healthy persons, CHB patients with HBV-DNA negative (20 cases) and HBV-DNA positive (24 cases) for up to 72 h. Apoptosis and Fas expression of the HK-2 were detected by flow cytometer. RESULTS: The apoptosis rate and Fas expression of HK-2 cells were significantly higher in HBV DNA positive serum group 19.01%+/-5.85% and 17.58%+/-8.35%, HBV DNA negative serum group 8.12%+/-2.80% and 6.96%+/-2.76% than those in control group 4.25%+/-0.65% and 2.33%+/-1.09%, respectively (P<0.01). The apoptosis rate and Fas expression of HK-2 in HBV DNA positive serum group was significantly higher than those in HBV DNA negative serum (P<0.01). Apoptosis rate of HK-2 cells in HBV DNA positive serum group was positively correlated with the level of HBV-DNA (r = 0.657). The level of serum TGF-beta (1) in CHB group was 163.05+/-91.35 microg/L, significantly higher as compared with 81.40+/-40.75 microg/L in the control group (P<0.01). CONCLUSION: The serum of patients with chronic hepatitis B promotes apoptotic damage in human renal tubular cells by triggering a pathway of Fas up-regulation. HBV and TGF-beta (1) may play important roles in the mechanism of hepatitis B virus associated glomerulonephritis.