Study On The Anti-HSV Activity Of Effective Parts From Duchesnea India(andr.) Focke Extracts

Objective: Observing the effect of Duchesnea india(Andr.) Focke against herpes simplex virus, influenza virus, respiratory syncytial virus, coxsackie virus and Enterovirus71 in vitro. screening the sensitive virus to extract and isolate the effective ingredients which has high efficient antiviral actions in vitro from the Duchesnea india(Andr.) Focke. Verify the antiviral effect of effective ingredients on viral animal model in vivo. Explore the antiviral mechanism of effective ingredients.Methods: Using neutral red staining and observation of cytopathic effect(CPE) can detect Duchesnea india(Andr.) Focke extracts for inactivation of virus in vitro, and therapeutic index(TI) was taken as indexes to screen the sensitive viruses to the drug; The water extract of Duchesnea indica focke alcohol precipitation process optimization,to get the optimal alcohol precipitation technology; Three different types of macroporous adsorptive resins was used to purify the supernatant,we also used therapeutic index(TI) as indexes to optimize the optimum macroporous adsorptive resin for high efficient antiviral actions in vitro; The supernatant was absorbed by D101 macroporous resin, and then the resin was eluted with water, 25% ethanol and TLC separation with the index of therapeutic index(TI),to get antiviral effective part; Mouse model with herpes simplex virus was established to evaluate the antiviral efficacy in vivo; The establishment of 4 types of virus-infected cell model,explore the antiviral mechanism of effective part.Result: Screening antiviral spectrum experiment in vitro, Duchesneaindia(Andr.) Focke extract anti HSV-1 effect was most pronounced, its therapeutic index reached at 15.09; The best process of alcohol precipitation for drug ratio of solid to liquid 1:1.3,alcohol concentration of alcohol precipitation after the 60%, alcohol precipitation time 24h; Byusing three types of macroporous adsorption resin was carried out on the supernatant after separation, it is concluded that antiviral effect of D101 macroporous adsorption resin is better in vitro;Through macroporous adsorption resin and the preparation of thin layer isolated spots have significantly antiviral effect, its therapeutic index reached at 51.34;Hepatitis in mice model was established by HSV-1,take liver tissue of effective part group compared with virus group, liver tissue can be observed in lesions less from the appearance, through the analysis of animal liver tissue pathology index, effective part group compared with virus model group have significant difference(P=0.027<0.05); The anti-HSV-1 mechanism of Spot position found that the effective parts of virus without direct killing effect, but the virus of wear into effect, reduce the combination of cells and viruses,it has influenced a functional protein of HSV-1, suppressing the virus reverse transcription and replication process.Conclusion: This experiment takes antiviral treatment index(TI) as the evaluation index in vitro, to phase separation and purification of antiviral activity parts, the study found spots part can can be suppressed mice hepatic tissue infection caused by HSV-1, and shows that it has more significant antiviral effect in vivo. The effective parts can prevent the virus, and suppress the virus biosynthesis, but no direct killing effect.