title: Identification and evaluation of anti hepatitis C virus phytochemicals from Eclipta alba.
abstract: BACKGROUND
  Eclipta alba, traditionally known as bhringraj, has been used in Ayurvedic medicine for more than 1000 years in India. It is used for the treatment of infective hepatitis, liver cirrhosis, liver enlargement and other ailments of liver and gall bladder in India. The aim of this study was to evaluate anti-hepatitis C virus activity present in the Eclipta alba extract, perform bioassay based fractionation and identify anti-HCV phytochemicals from the active fractions.
  
  
  METHODS
  Identification of active compounds was performed by bio-activity guided fractionation approach. Active isolates were separated by the combination of silica gel chromatography and preparative scale reverse phase HPLC. Eclipta alba extract and its isolates were examined for their ability to inhibit HCV replicase (HCV NS5B) activity in vitro and HCV replication in a cell culture system carrying replicating HCV subgenomic RNA replicon. The purified isolates were also examined for their binding affinity to HCV replicase by fluorescence quenching and their cytotoxicity by MTT assay.
  
  
  RESULTS
  Eclipta alba extract strongly inhibited RNA dependent RNA polymerase (RdRp) activity of HCV replicase in vitro. In cell culture system, it effectively inhibited HCV replication which resulted in reduced HCV RNA titer and translation level of viral proteins. Bioassay-based fractionations of the extracts and purification of anti-HCV phytochemicals present in the active fractions have identified three compounds, wedelolactone, luteolin, and apigenin. These compounds exhibited dose dependent inhibition of HCV replicase in vitro, and anti-HCV replication activity in the cell culture system
  
  
  CONCLUSIONS
  Eclipta alba extract and phytochemicals isolated from active fractions display anti-HCV activity in vitro and in cell culture system. The standardized Eclipta alba extract or its isolates can be used as an effective alternative and complementary treatment against HCV.
authors.affiliation: Department of Biochemistry and Molecular Biology, Center for Emerging Pathogens, UMDNJ-New Jersey Medical School, 185 South Orange Avenue, Newark, NJ 07103, USA.
authors.forename: Dinesh
authors.initials: D
authors.lastname: Manvar
authors.forename: Mahesh
authors.initials: M
authors.lastname: Mishra
authors.forename: Suriender
authors.initials: S
authors.lastname: Kumar
authors.forename: Virendra N
authors.initials: VN
authors.lastname: Pandey
mesh_terms: D000998:Antiviral Agents / Q000302:isolation & purification / Q000494:pharmacology*; D045744:Cell Line, Tumor; D036601:Eclipta; D016174:Hepacivirus / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D010936:Plant Extracts / Q000494:pharmacology*; D012367:RNA, Viral / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D010936:Plant Extracts; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase