Antiviral Effects Of EGCG Palmitate On Hepatitis C Virus In Vitro And Mechanism Study

Human,as an important members on earth,are changed by the environmental when people changes their survival environment. Though destroying the internal environment homeostasis of human, pathogens lead to many diseases.Hepatitis C virus (HCV) is a kind of global distributed single-stranded RNA viruses, according to the variation region sequence of its genome,HCVcan be divided into six genotypes as well as hundreds of species of subtypes. After infection in humans, the growing virus can change normal ecological environment in cells through a long period of incubation period, so that chronic hepatitis gradually develop in cirrhosis, liver cancer or fulminant hepatic failure resulting in liver function completely lost, eventually claiming the lives of those infected.This paper aims at studying the tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) palmitate impact on HCV in vitro and try to illustrate the interaction betweenthe EGCG palmitate and HCV and Huh7.5.1 cells by using Molecular Biology detection means from the micro ecology point of perspective.Research at the beginning, using CCK-8 kit to detect toxic effects of EGCG palmitate on Huh7.5.1 Cells, measuring HCV titer by Hepatitis c virus nucleic acid quantitative detection kitusing real-time PCR to selected the optimal multiple of infection(MOI) of infection, and then study its drug antiviral effects.Detecting the phosphorylation level of cellular interferon pathway STAT2 the role of EGCG palmitate by western blotting. Through RNA interference detection of the heat shock protein (HSPA5) to confirmed the relationship between its gene expression and EGCG palmitate antiviral effect with HCV.The results:0～50μg/mL of EGCG palmitate is harmless to Huh7.5.1, efficient to prevent HCV into cell and reproduction., but the effect of protective does not charge with adding EGCG palmitate concentration.The concentration of drug was increased, the effect of inhibit viral multiplication is enhanced. With 25 μg/mL the inhibition effect of drug is the best,the gene expression of virul is 1/6 of positive control group.Dealing cell with continue low leverl of drug, the proliferation of virus can be reduced large. At the same time, the groud, handle with IFN-a,viruls gene expression is half of negative groud, proved the phenomenon that interferons alpha cure HCV patients treatment less than 50% effectiveness. We hypothesized that EGCG palmitate can supersede interferons alpha becoming efficient therapeutic drug.Through the western bolting experiment we found that phosphorylation modification STAT2 is critical for EGCG palmitate tantiviral activity, the antiviral effects of EGCG palmitate was reduced when the expression of HSPA5 protein was interfered by lentivirus packaging. Though the experimental result, the gene expression of virul was reduced when expression of HSPA5 interfered, we knowed that HSPA5 plays an important host factor of HCV infection, the antiviral effects of EGCG palmitate may be realized by phosphorylation modification STAT2,up-regulation of antiviral gene expression to inhibiting proliferation of vims. Meanwhile, specificity combined with HSPA5 to reduce the activity of protein to control the virus proliferation direct. The antiviral effect of EGCG palmitate is achieved though two ways working together, of course, the environment of cell survival is very micro and precise, many other ways to come ture EGCG palmitate’s antiviral effect intracellular. Hope that this study can contribute to overcome HCV.