Inhibitory Effect Of Garlic Polysaccharide On EB Virus EA,VCA Expression And DNA Replication In Vitro

Objective: To discuss the effects of garlic polysaccharide(GPs) to expressions of EB virus early antigen(EA) and capsid antigen(VCA) and its DNA replication in vitro.Methods: Adopting cck-8 assay detected GPs' safe concentrations level which has no effects on cell activity. After EA in Raji cells and VCA in B95-8 cells were induced by sodium butyrate(Na B, 4m M/m L) and croton oil(CTO, 500 ng/ml), we detected the effects of GPs on the expressions of EB virus EA and VCA by indirect immunofluorescence(IIF). And the effects of GPs on the expression of EB virus transcription factor BZLF1 in lytic cycle replication was detected by q RT-PCR assay. Meanwhile, using FCM assay to detect the effects of GPs' safe concentrations level on Raji cells apoptosis and indirectly affirm the effects on cell activity. Raji cells for stage I EB virus latent infections can be induced by Na B/CTO into lytic cycle replication, thus we detected the effects of GPs on the EB virus DNA by q PCR assay and EBNA-1 m RNA by q RT-PCR assay.Result: 1. 2.00mg/ml or less GPs for the safe concentrations of Raji and B95-8 cells, and the concentrations in 24, 48 and 72 h is no difference(P > 0.05).2. Compared with the positive control, 1mg/ml GPs could inhibit EA antigen expression(12.55%) and 2mg/ml(62.70%) in Raji cells that induce-expressed EA, the difference statistically significant(P < 0.05).3. Compared with the positive control, 1mg/ml GPs could inhibit VCA antigen expression(43.86%) and 2mg/ml(66.92%) in B95-8 cells that induce-expressed VCA, the difference statistically significant(P < 0.05)4. The BZLF1,a key transcription factor for Initializing lytic replication to EBV, was detected by q RT-PCR in Raji cells that induce-expressed EA. The results show the inhibition levels of BZLF1 were 4.86%(P > 0.05) and 19.44(P < 0.05) in 1.00 and 2.00mg/ml GPs, respectively.5. The FCM results show the safe concentrations of GPs for Raji cells apoptosis have no differences between treatment and control group(P > 0.05), The above results show that GPs on both EA and VCA has obvious inhibitory effect in vitro, are dose dependent.6. The results of GPs anti-EBV replication show GPs inhibitions rates of EB virus DNA is 38.7% for latent replication and lytic replication is 39.06%. And the EBNA-1 m RNA were 38.7% and 36.4% in latent and lytic replication, respectively. The above results show that GPs for EB virus replication has obvious inhibitory effect in vitro(P < 0.05).Conclusion: 1. the GPs safe concentration of 2.00 mg/ml or less in both Raji and B95-8 cells.2. GPs could inhibits the EB virus EA and VCA expression and DNA replication in vitro, and it are dose dependent.