ABSTRACT
Ohjective To observe the inhibition effect of total glucosides of Picrorhiza on hepatitis B virus covalently closed circular DNA (HBV cccDNA) in HepG 2.2.15 cell line. Methods HepG 2.2.15 cells were incubated with culture medium containing 50 mg/L of picrosides or 5 mg/L of adefovir dipivoxil for 2 or 5 days. HBV DNA in the supernatant, intracellular cccDNA, relaxed circular DNA (rcDNA) and pregenomic RNA (pgRNA) were quantified by specific real-time polymerase chain reaction (RT-PCR) and inhibition rates were calculated. The means were compared by t test. Results After treated with picrosides for 2 and 5 days, the inhibition rates of HBV DNA in thesupernatant were 49. 74% (t=4.723, P<0.05) and 79.48% (t = 7.512, P<0.05), respectively. The inhibition rates of intracellular cccDNA were 43.55% (t = 5.216, P<0.05) and 56.43% (t=7.262, P<0.05), respectively, while those of intracellular rcDNA were 43.39% (t=4.137, P<0.05) and 63.86% (t=7.861, P<0.05), respectively, and those of intracellular pgRNA were 54.72% (t=4.532, P<0.05) and 56.08% (t=4.833, P<0.05), respectively. Comparatively, after treatment with adefovir dipivoxil for 2 and 5 days, the inhibition rates of HBV DNA in the supernatant were 25.56% (t=2.874, P<0.05) and 92.44% (t =10.276, P<0.05), respectively. Those of cccDNA were 18.54% (t=2.736, P<0.05) and 47.19% (t=6.852, P<0.05), respectively. Those of rcDNA were 21. 20% (t=3.206, P<0.05) and 71.47% (t=8.332, P<0.05), respectively, pgRNA were 11.14% (t=1.761, P>0.05) and 37.61%(t=3.632, P<0.05) respectively in HepG2.2.15 cells. Conclusions Pierosides may inhibit the replication cycle of HBV, including the formation of cccDNA in HepG 2.2.15 cells. The mechanism of pierosides on cccDNA may differ from adefovir dipivoxil's due to its earlier inhibition time phase.
ABSTRACT
Objective To understand the etiology of acute hepatitis B (AHB) in adults and investigate the mechanisms of hepatic injury and viral clearance in AHB. Methods One hundred and twenty adult AHB patients were enrolled. Epidemiological and clinical data were collected from the case history records or face-to-face inquiry, and serum samples were collected during hospitalization and follow-up. To observe dynamic patterns of AHB etiology, the markers of hepatitis B virus (HBV) were detected by enzyme-linked immunosorbent assay (ELISA); the level of HBV DNA and HBV genotype were determined by real-time polymerase chain reaction (PCR). Enumeration data were analyzed by non-parametric rank sum test. Comparison between groups was done by t test and that between rates of samples was done by Pearson χ2 test. Results Serum HBV DNA was positive in 48.33% of patients at the time of diagnosis with mean level of 9.84×04 copy/mL, and became undetectable after 12.5 days on average. The median levels of serum alanine aminotransferase (ALT) were 1600 U/L and 1490 U/L in HBV DNA positive and negative groups, respectively (z=-0. 678, P=0. 498). However, the mean levels of serum ALT were (2058±123) U/L and (1393±139) U/L in groups of HBV DNA<1×104 copy/mL and>1×104 copy/mL, respectively, which was significantly different (t=-2.17, P=0. 049). Genotype B accounted for 52.5%, genotype C 42.5 and genotype B and C mixed type 5.0% in 58 patients with HBV DNA positive. Eight patterns of serum HBV markers were presented at first visiting. HBsAg(+), HBeAg(+), anti-HBc(+), anti-HBc IgM(+) and HBsAg(+), anti-HBe(+), anti-HBc(+), anti-HBc IgM(+) were the most common patterns, which accounted for 38.3% and 30.0%, respectively. The dynamic patterns of serum HBV markers of 28 AFIB patients were prospectively followed up. The rate of serum FIBsAg loss was 100. 0% and the median time of negative-conversion was 3 weeks. The cumulative positive rate of anti-HBs was 85.7% after 52 weeks of follow-up. The rate of serum HBeAg loss was 100.0%. HBeAg was negative in 53.6% of patients at first visiting and the rest of patients achieved negative within 4 weeks after onset. The positive rate of anti-HBe was 82.1% during 52 weeks of follow-up. Total anti-HBc (including IgG and IgM) was keeping positive in all patients within 52 weeks, and the negative rate of anti-HBc IgM was 39. 3% after followed up for 52 weeks. Conclusions Rapid HBV clearance andserum HBV marker conversion are significantly different between AHB and chronic hepatitis B.