| Hepatitis B is a liver disease that can cause a variety ofcomplications. Hepatitis B virus (HBV) infection is one of the major publichealth problems in the world. China is a high-prevalence area for HBVinfection. The number of HBV carriers in china is up to10%of the totalpopulation. Of these, approximately30million people are the patients withchronic hepatitis B, Moreover, about35million people every year died fromchronic hepatitis B-related diseases. However, no drug or therapeutic methodcan cure this disease so far. There is still a very long distance from the efficacyof the existing anti-hepatitis B drugs to the goal to cure the vast majority ofthe patients with HBV. Therefore, it is of great significance to carry out theresearch on the potential targets and new drugs for anti-HBV treatment.1. Induction of heme oxygenase-1expression and its anti-HBV propertyHemin was dissolved with ammonia and ultra-pure wate. HepG2.2.15cells were treated with the different concentrations of hemin for1-6days. Todetect the expression of HO-1gene with reverse transcription PCR (RT-PCR)method, total RNA of HepG2.2.15cells was extracted and then reversetranscribed into complementary DNA (cDNA). β-actin was used as an internalcontrol. The levels of HBsAg and HBeAg in replaced culture supernatantswere respectively determined by enzyme-immunoassay kits. The HBV viralload in replaced culture supernatants was detected with a HBV DNAPCR-fluorescence quantitation kit. The results showed that hemin at5-100μM induced significantly the expression of HO-1. With the increasing of HO-1expression, the HBsAg/HBeAg secretion and HBV-DNA replication wereinhibited accordingly. Furthermore, with the extention of the interaction timeof hemin and HepG2.2.15cells, the inhibition trend was more obvious.2. Effect of anti-HBV of cichoric acid and other natural extracts.HepG2.2.15cells and D-galactosamine (D-GalN)-induced HL-7702cellinjury model were employed to assess the anti-HBV and hepatocyteprotective effects of three natural extracts (including cichoric acid, T. mongolicum and L.alata extracts), respectively. The cytotoxicity of test extracts at the differentconcentrations on HepG2.2.15cells and HL-7702cells were respectivelydetermined by the MTT assay. HepG2.2.15cells were treated with thedifferent concentrations of test drugs and lamivudine for6days. The replacedmedium was detected for the measurement of the HBsAg/HBeAg andHBV-DNA levels with enzyme-linked immunosorbent and fluorescencequantitative PCR assay, respectively. The results showed that test drugs at thedifferent concentrations improved markedly HL-7702cell viability and theirhepatoprotective effects were stronger than the positive control drug silibininunder the same concentration. Test drugs exhibited the significant inhibitionon the HBsAg/HBeAg secretion and HBV-DNA replication in HepG2.2.15cells and had a dose-and time-dependent manner. Furthermore, test drugsafforded stronger anti-HBV effects than that of the positive controllamivudine.In summary, we studied the overexpression of HO-1induced by hemin inHepG2.2.15cells and discussed the relation of HBV replication (includingHBsAg/HBeAg expression and HBV DNA replication) and induction of HO-1.The study verifies the direct anti-HBV effect of HO-1induction and providesthe partial scientific evidence of HO-1as a potential target for the treatment ofhepatitis B and development of anti-HBV drugs. In addition, the anti-HBVand hepatocyteprotective effects of three natural extracts were evaluated bythe HepG2.2.15cells and HL-7702cell injury model induced by D-GalN,respectively. These data laid a foundation for the development and utilizationof natural phenolic acids, especially caffeoyl analogues. |
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