title: 18β-glycyrrhetinic acid inhibits rotavirus replication in culture.
abstract: BACKGROUND
  Glycyrrhizin (GA) and primary metabolite 18β-glycyrrhetinic acid (GRA) are pharmacologically active components of the medicinal licorice root, and both have been shown to have antiviral and immunomodulatory properties. Although these properties are well established, the mechanisms of action are not completely understood. In this study, GA and GRA were tested for the ability to inhibit rotavirus replication in cell culture, toward a long term goal of discovering natural compounds that may complement existing vaccines.
  
  
  METHODS
  Epithelial cells were treated with GA or GRA various times pre- or post-infection and virus yields were measured by immunofluorescent focus assay. Levels of viral proteins VP2, VP6, and NSP2 in GRA treated cells were measured by immunoblot to determine if there was an effect of GRA treatment on the accumulation of viral protein.
  
  
  RESULTS
  GRA treatment reduced rotavirus yields by 99% when added to infected cultures post-- virus adsorption, whereas virus yields in GA treated cultures were similar to mock treated controls. Time of addition experiments indicated that GRA-mediated replication inhibition likely occurs at a step or steps subsequent to virus entry. The amounts of VP2, VP6 and NSP2 were substantially reduced when GRA was added to cultures up to two hours post-entry.
  
  
  CONCLUSIONS
  GRA, but not GA, has significant antiviral activity against rotavirus replication in vitro, and studies to determine whether GRA attenuates rotavirus replication in vivo are underway.
authors.affiliation: Immunology and Infectious Diseases, Montana State University, Bozeman, MT 59718, USA. mhardy@montana.edu
authors.forename: Michele E
authors.initials: ME
authors.lastname: Hardy
authors.forename: Jay M
authors.initials: JM
authors.lastname: Hendricks
authors.forename: Jeana M
authors.initials: JM
authors.lastname: Paulson
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Faunce
mesh_terms: D000818:Animals; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006034:Glycyrrhetinic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology; D019695:Glycyrrhizic Acid / Q000494:pharmacology; D000882:Haplorhini; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D012401:Rotavirus / Q000187:drug effects* / Q000502:physiology*; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D016601:RNA-Binding Proteins; C442545:VP2 protein, Rotavirus; C061332:VP6 protein, Rotavirus; D017361:Viral Nonstructural Proteins; C079234:NS35 protein, rotavirus; C119129:18alpha-glycyrrhetinic acid; D019695:Glycyrrhizic Acid; D006034:Glycyrrhetinic Acid