The Study Of Lamivudine Combined With Protocatechuic Acid Against Duck Hepatitis B Virus In Vivo

Hepatitis B virus (hepatitis B virus, HBV) infection is a worldwide problem, nearly2billion people infected by the hepatitis B virus (HBV). According to WHO reports, it will not only cause acute, chronic hepatitis, cirrhosis of the liver and cancer. About350million chronic HBV infected persons who died from liver failure, and about1million person died from cirrhosis and hepatocellular carcinoma (HCC) is. With statistics, in China, about12million patients are infected by Hepatitis B virus, and120million people carried HBsAg, with1/3of global hepatitis B virus carriers.Lamivudine (lamivudine,3TC) is the current first-line drugs to treat hepatitis B. It caused a significant inhibition on HBV-DNA with hepatitis B patients, but the long-term efficacy of drugs, anti jump high, and more side effects, may easily cause a mutation of the virus and other problems to limit its clinical application. Our previous studies showed that compared with the3TC single time, the PA and3TC in combination had synergistic inhibitory effect on secretion of HBV DNA and antigens in HepG2.2.15cells, DHBV adsorption, HBV X protein into the nucleus, entry and infection of duck primary hepatocytes.At the same time, the PA and3TC in combination caused significant pharmacokinetic interaction:slowed3TC absorption process and increase total absorption; Slowed PA absorption rate and elimination, extended its role time in vivo. In this experiment, We took ducklings infected with duck hepatitis B virus (DHBV) as experimental animals in the pre-optimized on the basis of the PA and3TC combination dose ratio, to further investigated its dose-effect relationship and time-effect relationship for anti-viral and protecting liver in vivo, in order to develop the experimental foundation of new anti-HBV drug combinations-3TC/PA.Methods:(1) the amount of-activity relationship experiments:five doses3TC/PA (12.5/12.5mg/kg,25/25mg/kg,50/50mg/kg,100/100mg/kg,150/150mg/kg) Eachdays, respectively, gavage administration for14days.Respectively before injection3days after administration of7days,14days and withdrawal blood serum of DHBV of DNA content of the albumin (ALB) and total bilirubin (Tbil) and aspartate aminotransferase (GOT), alanine transaminase (GPT) and alkaline phosphatase (AKP) activity.(2)-effect relationship experiments:3TC/PA (50/50mg/kg) daily gavage once a continuous administration of28days.Prior to administration and5days after the administration of the first7days,14days,21days,28days and withdrawal blood serum DHBV DNA、DHBsAg、DHBcAbcontent, and the GOT and GPT activity.Results:1. Experiments of Dose-effect relationship:the studies have shown that the inhibitory effect of3TC/PA for DHBV DNA in serum is increased with dose (12.5-150mg/kg) increases, but when stopped lavage for3days, in the high dose (100-150mg/kg)sample, the levels of virus DNA appeared rebound. During the administration and3days after stopping, in the dose group of50mg/kg3TC/PA, the content of viral DNA is significantly lower than the acyclovir control group of100mg/kg,and the GOT, GPT, and Tbil value were significantly lower than the door bird group of5g/kg.2. As time-effect studies shown, not only the inhibitory effect of50/50mg/kg3TC/PA for DHBV DNA, DHBsAg, GPT and GOT in serum is strengthened with extension of the dosing cycle during the0-28d administration period, and the levels of viral DNA,DHBsAg in serum is significantly lower than the acyclovir control group of100mg/kg during the administration and5day after stopping,And also the GPT and GOT levels in serum were significantly lower than the door bird control group of5g/kg.Conclusion:the most appropriate anti-DHBV dose in vivo is50/50mg/kg3TC/PA for ducklings infected with DHBV. And the effect of anti-viral is better than acyclovir of100mg/kg, the effect of protecting Liver is better than the door birds of5g/kg.