The Molecular Mechanism Of EGCG-p Anti-EV71 Virus

Enterovirus EV71 is the most important virus causes Hand-foot-mouth disease (HFMD). There are millions of children suffering from HFMD every year. However there is no effective drug treatment for it. EGCG is a kind of natural antiviral drugs, it has been proven to have a variety of virus antiviral effect. EGCG has an effect on resistance to multiple viruses, and its ingredients is derived from tea with little side-effect, which has enormous potential for developing new drugs. EGCG is not stable in the water, and easily oxidized leading to losses. Therefore, we used the modified lipid soluble EGCG-p which have been proven to have antiviral activity that combined with fatty acid palmitate, EGCG-p improves the stability of compounds. So far, few people make a study on EGCG-p anti EV71, EGCG-p anti EV71 is still an unknown field. In order to verify the effectiveness of EGCG-p anti EV71, this paper made a preliminary study on EGCG-p vitro anti-EV71, it aims to provide the theoretical basis and experimental basis for the further development of clinical applications.EV71 proliferative activity in human rhabdomyoma cells (RD cells) cell is more active than others, proliferation efficiency is higher than others, so we do in vitro experiments on RD cell as a cell matrix. In order to study the capacity of EGCG in antiviral activity of EV71, this study used three drug delivery procedure(adding drugs,then adding virus; adding virus, then adding drugs; virus and drugs at the same time to the RD cells) to study EGCG against EV71 virus. We first tested the virulence of EV71 virus, then used different orders to add virus and drugs to experiment. When cells have obvious lesions, we used chloroform extraction RNA method to extract RNA, and then reversed to get the EV71 virus cDNA, and finally carried on the fluorescent quantitative PCR detection. The result shows that adding drugs first then virus has significant effect. Accordingly, we think that EGCG-p has the effect of preventing EV71 virus.In order to further study on molecular mechanism of EGCG-p against EV71 virus, this research cloned and expressed SCARB2 receptor protein and EV71 virus protein VP1, used molecular interaction analyzer for SCARB2, EGCG-p to interact with VP1.SCARB2 receptor and VP1 were tested, the results showed that the affinity average of interaction between SCARB2 receptor and VP1 is 0.045μM, Full R2 is 0.96,indicating that the software fit calculation reliability is high. Then we made that SCARB2 receptors interact with EGCG-p, calculated the affinity average of interaction between SCARB2 receptor and EGCG-p, the affinity average is 0.0176μM, Full R2 is 0.95,indicating that the software fit calculation reliability is high. At last, we made that SCARB2 plus EGCG-p interact with VP1. the affinity average of interaction between SCARB2 receptor and VP1 is 0.045μM.The affinity average of interaction between SCARB2 plus EGCG-p VP1 is 0.549μM, Full R2 is 0.925.It is concluded that in the presence of EGCG-p, the affinity of SCARB2 and VP1 decreased, from the molecular level, indicating that EGCG-p has antiviral effect.From these results it can be concluded that EGCG-p can combine with receptor proteins SCARB2, stop the invasion of EV71 virus, from molecular level proving the EGCG-p has antiviral effect.