| Summary: | 碩士 === 中國醫藥大學 === 醫學檢驗生物技術學系碩士班 === 99 === Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus with positive-sense single-stranded RNA genome. JEV infection is asymptomatic, only few cases have headache, fever, encephalitis, coma and even death. Treatment for JE is supportive and still lack of efficacy. Water extrats of Pericarpium granati (PG) is an traditional herbal therapeutic with antiviral activities. It was shown to inhibit influenza A virus replication in chicken embryos and herpes simplex viruses in cell culture. In this study, we intend to investigate the anti-JEV activities and functional mechanism of PG water extract and the related compound punicalagin (PN). CC50 values (50% cytotoxicity concentration) of PG water extract by MTT test were 53 μg/mL for BHK-21 cells and 219.7 μg/mL for TE-671 cells, while CC50 values of PN were14 μM for BHK-21 cells and 42.7 μM for TE-671 cells. PG water extract and PN showed a concentration-dependently inhibitory effect on JEV-infected cytophatic effect and virus production in BHK-21 and TE-671 cells. The plaque reduction abilities of PG water extract and PN were studied by three different procedures including addition at 1 hour before virus inoculation (pre-treatment), addition and virus inoculation at the same time (simultaneous treatment), and addition at 1 hour after virus inoculation (post-treament). IC50 values (50% inhibition concentration) of PG water extract were 4.4 μg/mL by pre-treatment, 5.2 μg/mL by simutaneous treatment and 74.6 μg/mL by post-treament. IC50 values (50% inhibition concentration) of PN were 3.27 μM by pre-treatment, 1.17 μM by simutaneous treatment and 9.18 μM by post-treament. In vivo signaling pathway assay indicated PG water extract and PN inducing activation of the NF-κB signaling pathway.
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