| Hepatitis C virus (HCV) infection has become a world health problem, affecting millions of people in the world. Most infection of HCV become persistent, and many chronically infected individuals will develop liver cirrhosis, fibrosis and even hepatocellular carcinoma (HCC). The variability of HCV genome and lack of cell culture system and small animal model supporting HCV production are the major impediments in HCV research and anti-HCV drug and vaccine development.In this study, according to the structure of genotype lb HCV genome clone, the high level of HCV RNA replication was detected in the non-liver cell line Vero, helped with the T7 RNA polymerse provided by recombinant vaccinia virus vTF7-3. HCV positive and negative RNA both synthesized in the transfected Vero cells. Western blot analysis indicated the expression of HCV envelope proteins increased with the time increase of transfection. According to the results, we chose the best time to collect virus. After purification, the HCV virions with a diameter of 50nm approximately were observed under the electron microscopy.The HCV virions (HCVcc) expanded from Vero cell culture system transfected with full-length HCV genome infected HCV host cell lines, and the results showed that HCV RNA could be effectively synthesized and proteins expressed in the host cells, and the supernate of infected cells was also infectious.Since the chimpanzee is the only nonhuman host serving as a model for HCV infection and the availability of these animals is limited, the development of new model systems is pivotal for the analysis of HCV infection. The tree shrew Tupaia belangeri a species closely related to primates, has been shown to be susceptible to a variety or human viruses, including herpers smiples, hepatitis B virus and rotavirus. The results demonstrated that tree shrew could be efficiently infected with HCVcc, and appeared viremia and pathological changes in liver like as the symptom of infected people. The establishment of this small animal model will advance the study of HCV infection and anti-HCV drug.HCV infection is a major cause of liver disease. The current standard in hepatitis C treatment schemes consistingin combination regiments of pegylated interferon-a with Ribavirin are quite successful in patients with HCV genotypes 2 and 3 infections achieving HCV eradication rates of 75-90%. However, they are much less effective in patients with genotypes 1 and 4 infections. Morever, they have several adverse effects and contraindications, further limiting their efficacy and applicability in an appreciable number of patients with chronic HCV-induced liver disease. Therefore, the need for improvement of existing therapies and for development of new effective, safe and tolerable drugs is a matter of great clinical relevance and importance.Antimicrobial peptides (AMPs), rich sources existing in nature, are able to effectively kill multidrug-resistant pathogens. Scorpion venom is a rich resource of AMPs, and scorpion venomous peptides have benn reported to have the antiviral activity. In this study, we screened 6 clones from the cDNA library of the venomous gland having antimicrobial activity. But HCV RNA inhibitory assays indicated that only Hp1090 showed well antivirus activity, and prevents initiation of HCV infection. Futhermore, Hp1090 was detected could interact with HCV particles indirectly and permeabilize phospholipid membranes rapidly. These results suggested that Hp1090 could be considered a potential anti-HCV drug and the action of destabilizing the lipid composition of viral membranes offer a unique therapeutic approach to HCV infection. |
PDF Full Text Request