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Title:	Antiviral action of Phaleria macrocarpa fruits against herpes simplex virus type-1 (HSV-1)
Authors:	Mahmud Yusef Yusef Ismaeel (P72867)
Supervisor:	Nazlina Ibrahim, Assoc. Prof. Dr.
Keywords:	Phaleria macrocarpa Herpes simplex virus Antiviral action Fruit extracts Mahkota Dewa Dissertations, Academic -- Malaysia
Issue Date:	10-Feb-2016
Description:	This study aims to determine the antiviral activity of Phaleria macrocarpa (Mahkota Dewa) fruit against herpes simplex virus type-1 (HSV-1). Dried fruits were extracted using methanol and water to produce methanolic extract (ME). Further fractionation produce hexane fraction (HF), chloroform fraction (CF) and ethyl acetate fraction (EAF). The fruit of P. macrocarpa was also extracted with water to produce aqueous extract (AE) and freeze dried. Qualitative phytochemical analyses revealed the presence of steroids and flavones aglycones in the ME, HF, CF, EAF and AE. Terpenes, alkaloids, saponins and tannins were present in ME, EAF and AE but not in CF. Terpenes were present in HF but saponins, alkaloids and tannins were absent. The cytotoxicity towards Vero cell was evaluated using MTT assay with 50% cytotoxic concentration (CC50) value ranges between 0.40 to 5 mg/mL. The findings indicate that AE, ME and the fractions are safe and not toxic towards Vero cells. Screening by plaque reduction assay showed that AE, ME and its fractions except CF have antiviral activity against HSV-1. The selectivity indices (SI) ranged between 2.6 and 17.9. From the SI values it is worth for further investigation to determine the antiviral mode of action. In the cell pretreatment assay, cells were treated and followed by infection with HSV-1. No plaques were formed in the assay when treated with either ME, AE or the fractions. This indicates that the extracts or fractions were able to halt virus infection. In the virucidal assay, AE, ME and its fractions showed potential antiviral property with the percentage of formation value ranges from 30.5% to 91.26% at 1×102 pfu/mL. In addition, plaque formation was completely inhibited when virion was treated with either AE, ME or fractions at 1×106 pfu/mL and 1×104 pfu/mL. The highest antiviral activity shown by EAF of P. macrocarpa fruits and further analyses on the mode of action were done. In the penetration assay, EAF affects virus infection leading to inhibition to plaque formation. The anti-HSV-1 activity of EAF disables virus attachment to Vero cell. Virus treated with EAF at the concentrations of 0.20 mg/mL, 0.15 mg/mL or 0.10 mg/mL followed by infection to Vero cells showed inhibition of 46%, 31% and 22% respectively. Thus, EAF inactivates the early steps of viral life cycle of HSV-1 through virucidal and inhibit attachment in a dose-dependent manner. The time addition assay when treated with EAF at different time points maximum plaque inhibitory activity was 84% at 2 hours post infection (hpi) and descended gradually from 4 to 8 hpi. This indicated that the fraction inhibited the immediate early phase of the virus life cycle. In time removal assay, EAF antiviral activity was observed as early as 12 hpi and reached greater than 84% inhibition at 48 hpi. In conclusion, P. macrocarpa fruit extracts and fractions causes changes to cell which disables the ability of virus to infect and to virus in the capability to infect during attachment, penetration, immediate early replication and release.,Certification of Master's/Doctoral Thesis" is not available
Pages:	107
Publisher:	UKM, Bangi
Appears in Collections:	Faculty of Science and Technology / Fakulti Sains dan Teknologi

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