Studies On The Isolation And Purification Of Melittin And Its Effect On Anti-HSV-1 In Vivo And In Vitro

HSV-1 dued to the rising incidence of the disease and occured resistant strains of a virus in recent years. So we carried out the research on the anti-HSV-1 effect of melittin. The results were as follows:The venom of Apis mellifera lijustica was collected by QF-1 Super Auto Electronic BV-Collector and was isolated through polydextran gel. The separated component was approved to be at electrophoretic purification by electrophoresis. We needed store it for the following experiments.The anti-HSV-1 activity of melittin and ACV were evaluated by inhibition of viral cyctopathic effects. The results showed that 50 % toxic concentration of melittin (CC50) was 10-2.1 mg/100μL by MTT method. The viral suppression rate could reach 66.51 %, when melittin of 100μL 9CC50 was used. Fifty percent tissue culture infective dose (TCID50) is 10-5.2/100μL. Mice encephalitis model was established by infected HSV-1 respectively to evaluate the anti-HSV-1 efficacy in vivo. The results showed that the mice median lethal dose (LD50) was 10-3 mg/kg. Melittin increased the survival rate of mice model infected by HSV-1 from 20 % to 50 %, and inhibited HSV-1 obviously. Melittin can prolong the survival time of mice.The study was designed four groups. It would affect melittin on the direct effect of HSV-1, the viral adsorption, synthesis and release. The results showed: (1) Four concentration gradient of melittin (1CC50, 3CC50, 6CC50 and 9CC50) had an inhibition effect of adsorption of HSV-1. Their inhibition rates were 31.25 %, 32.92 %, 32.92 % and 32.08 %. And the highest inhibition rate was 32.92 %; (2) Four concentration gradient of melittin (1CC50, 3CC50, 6CC50 and 9CC50) had an inhibition effect of synthesis of HSV-1. Their inhibition rates were 29.05 %, 29.91 %, 31.69 % and 33.06 %. And the highest inhibition rate was 33.06 %; (3) Four concentration gradient of melittin (1CC50, 3CC50, 6CC50 and 9CC50) had an inhibition effect of release of HSV-1. Their inhibition rates were 29.31 %, 18.37 %, 13.28 % and 10.70 %. And the highest inhibition rate was 29.31 %. This showed that melittin could inhibit the release of HSV-1, but the inhibition rate of viral release was significantly reduced with increasing of melittin; (3) Direct effect of melittin on the viral experiment showed that test groups and the control group had the similar survival rate. It demonstrated that melittin had no direct role in killing the virus.