Proliferation Promoting And Protective Effect Of HYPER-IL-6 Recombinant Lentivirus On Hepatocytes And Rats Whith Acute Liver Failure

PARTâ… P ROLIFERATION PROMOTING AND PROTECTIVE EFFECT OF HYPER-IL-6 RECOMBINNANT ON HEPATOCYTESAIM: To investigate the proliferation promoting and protective effect of Hyper-IL-6 recombinant lentivirus on human normal hepatocytes L-02.METHODS: The constructed Hyper-interleukin-6 recombinant lentivirus (FIV-HIL-6), IL-6 recombinant lentivirus (FIV-IL-6) and empty lentivirus vector (FIV) were used to infect human hepatocytes L-02 respectively. The influences on cell cycle and proliferation cell nuclear antigen of hepatocytes L-02 were detected using flow cytometry. The content and expression changes of phospholipase A2 of hepatocytes L-02 infected by recombinant lentivirus were determined using ELISA and real-time quantitative fluorescence PCR.RESULTS: After virus particles infecting L-02 cells, the proliferation promoting effect of FIV-HIL-6 on L-02 cells was obviously higher than that in other control groups (p<0.01). In addition, the proliferation promoting effect on hepatocytes increased and reached the maximum at 72h after infecting. Compared with other control groups, the content of cytosolic phospholipase A2 of hepatocytes in FIV-HIV-6 group was obviously decreased (p<0.01). There was no obvious time effect on the influences of recombinant lentivirus infection on the content of phospholipase A2 of hepatocytes..Compared with hepatocytes in FIV group, the relative expression of phospholipase A2 mRNA of hepatocytes in FIV-HIV-6 group decreased.CONCLUSION: FIV-HIL-6 recombinant lentivirus has stronger proliferation promoting effect on hepatocytes L-02 and obviously reduces the content of phospholipase A2 of hepatocytes to further protect hepatocytes in inflammatory responses.PARTâ…¡PROTECTIVE EFFECT OF HYPER-IL-6 RECOMBINNANT ON RATS WITH ACUTE LIVER FAILUREObjective: To investigate the protective effect of recombinant lentivirus-mediated hyper-interleukin 6 (Hyper-IL-6) on rats with acute liver failure and its mechanism.Methods: Totally 45 rats with D-galactosamine acute liver failure were randomly divided into 3 groups: FIV-Hyper-IL-6 group, FIV-IL-6 group and non-infected groups (n=15), which were intraperitoneally injected with FIV-HIL-6, FIV-IL-6 and normal saline, respectively, at 2 h before modeling. At 12h, 24h, 48h, 72h and 7d after modeling, the liver function, endotoxin (ETM), lipid peroxidation levels (SOD, MDA) in liver tissue were determined. The light microscopy and electron microscopy were applied to observe histomorphological changes. The gene integration in liver tissue was detected by RT-PCR on the 7th day. Another 16 SD rats were randomly divided into 2 groups,which were given FIV-HIL-6, FIV peritoneal injection once, to observe the toxicity of recombinant lentivirus on rats.Results: In FIV-HIL-6 group, serum levels of alanine / grass transaminase (ALT/AST), total bilirubin (TB) value, MDA, ETM were lower than those in FIV-IL-6 group and non-infected group (P<0.05), while SOD value was higher. The recombinant lentivirus showed no significant toxic effects on rats. Hyper-IL-6 was expressed in liver tissue of rats.Conclusion: FIV-HIL-6 successfully transfected liver tissue of rats. For rats with acute liver failure,Hyper-IL-6 has strong anti-inflammatory and antioxidant effects, effectively improve liver function and histological condition,has protective and therapeutical effect effect on rats.