title: Effects of cocktail of four local Malaysian medicinal plants (Phyllanthus spp.) against dengue virus 2.
abstract: BACKGROUND
  The absence of commercialized vaccines and antiviral agents against dengue has made the disease a major health concern around the world. With the current dengue virus transmission rate and incidences, the development of antiviral drugs is of vital need. The aim of this project was to evaluate the possibility of developing a local medicinal plant, Phyllanthus as an anti-dengue agent.
  
  
  METHODS
  Cocktail (aqueous and methanolic) extracts were prepared from four species of Phyllanthus (P.amarus, P.niruri, P.urinaria, and P.watsonii) and their polyphenolic compounds were identified via HPLC and LC-MS/MS analysis. MTS assay was then carried out to determine the maximal non-toxic dose (MNTD) of the extracts, followed by screening of the in vitro antiviral activity of aqueous cocktail extracts against DENV2 by means of time-of-addition (pre-, simultaneous and post-) using RT-qPCR. The differentially expressed proteins in the treated and infected cells were analysed with two dimensional gel electrophoresis experiments.
  
  
  RESULTS
  Several active compounds including gallic acid, geraniin, syringin, and corilagen have been identified. The MNTD of both aqueous and methanolic extracts on Vero cells were 250.0 μg/ml and 15.63 μg/ml respectively. Phyllanthus showed strongest inhibitory activity against DENV2 with more than 90% of virus reduction in simultaneous treatment. Two-dimensional analysis revealed significantly altered levels of thirteen proteins, which were successfully identified by tandem MS (MS/MS). These altered proteins were involved in several biological processes, including viral entry, viral transcription and translation regulations, cytoskeletal assembly, and cellular metabolisms.
  
  
  CONCLUSIONS
  Phyllanthus could be potentially developed as an anti-DENV agent.
authors.forename: Sau Har
authors.initials: SH
authors.lastname: Lee
authors.forename: Yin Quan
authors.initials: YQ
authors.lastname: Tang
authors.forename: Anusyah
authors.initials: A
authors.lastname: Rathkrishnan
authors.forename: Seok Mui
authors.initials: SM
authors.lastname: Wang
authors.forename: Kien Chai
authors.initials: KC
authors.lastname: Ong
authors.forename: Rishya
authors.initials: R
authors.lastname: Manikam
authors.forename: Bobby Joe
authors.initials: BJ
authors.lastname: Payne
authors.forename: Indu Bala
authors.initials: IB
authors.lastname: Jaganath
authors.forename: Shamala Devi
authors.initials: SD
authors.lastname: Sekaran
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002522:Chlorocebus aethiops; D003715:Dengue / Q000821:virology*; D003716:Dengue Virus / Q000187:drug effects*; D005707:Gallic Acid / Q000494:pharmacology; D005960:Glucosides / Q000494:pharmacology; D006801:Humans; D047348:Hydrolyzable Tannins / Q000494:pharmacology; D010666:Phenylpropionates / Q000494:pharmacology; D029803:Phyllanthus / Q000737:chemistry*; D010936:Plant Extracts / Q000494:pharmacology*; D010946:Plants, Medicinal; D053719:Tandem Mass Spectrometry; D014709:Vero Cells
chemical_list: D000998:Antiviral Agents; D005960:Glucosides; D047348:Hydrolyzable Tannins; D010666:Phenylpropionates; D010936:Plant Extracts; C024603:Geraniin; D005707:Gallic Acid; C028305:syringin
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