title: A neutrophil multitarget functional bioassay to detect anti-inflammatory natural products.
abstract: A multitarget functional bioassay was optimized as a method for detecting substances interacting with the inflammatory process of activated neutrophil granulocytes, mainly to release elastase detected by p-nitroanilide (pNA) formation. Using this bioassay, 100 fractionated extracts of 96 plants were screened, with results presented in a manner that links recorded biological activity to phylogenetic information. The plants were selected to represent a major part of the angiosperms, with emphasis on medicinal plants, Swedish anti-inflammatory plants, and plants known to contain peptides. Of the tested extracts, 41% inhibited pNA formation more than 60%, and 3% stimulated formation. The extract of Digitalis purpurea enhanced pNA formation, and digitoxin, the active compound, was isolated and identified. Plant extracts that exhibited potent nonselective inhibition (>80% inhibition) were evaluated further for direct inhibition of isolated elastase and trypsin enzyme. The inhibitory effect of most tested extracts on the isolated enzyme elastase was similar to that of PAF- and fMLP-induced pNA formation. Compared to trypsin, inhibition of elastase by extracts of Rubus idaeus and Tabernaemontana dichotoma was significantly higher (80% and 99%, respectively). Inhibition of trypsin by the extract of Reseda luteola was high (97%). Orders such as Lamiales and Brassicales were shown to include a comparably high proportion of plants with inhibitory extracts.
authors.affiliation: Division of Pharmacognosy, Department of Medicinal Chemistry, Biomedical Centre, Uppsala University, PO Box 574, S-751 23 Uppsala, Sweden.
authors.forename: Senia
authors.initials: S
authors.lastname: Johansson
authors.forename: Ulf
authors.initials: U
authors.lastname: Göransson
authors.forename: Teus
authors.initials: T
authors.lastname: Luijendijk
authors.forename: Anders
authors.initials: A
authors.lastname: Backlund
authors.forename: Per
authors.initials: P
authors.lastname: Claeson
authors.forename: Lars
authors.initials: L
authors.lastname: Bohlin
mesh_terms: D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D001586:Benzoylarginine Nitroanilide / Q000032:analysis; D001681:Biological Assay / Q000379:methods*; D001688:Biological Products / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D003571:Cytochalasin B; D004070:Digitalis / Q000737:chemistry; D004072:Digitonin / Q000737:chemistry / Q000494:pharmacology; D004074:Digitoxin / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D004306:Dose-Response Relationship, Immunologic; D006461:Hemolysis; D006801:Humans; D019686:Lamiaceae / Q000737:chemistry; D019272:Leukocyte Elastase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000378:metabolism; D019684:Magnoliopsida / Q000737:chemistry*; D009240:N-Formylmethionine Leucyl-Phenylalanine; D018375:Neutrophil Activation / Q000187:drug effects; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology; D010455:Peptides; D010940:Plant Proteins / Q000032:analysis / Q000737:chemistry / Q000494:pharmacology; D010946:Plants, Medicinal / Q000737:chemistry* / Q000145:classification*; D010972:Platelet Activating Factor; D013481:Superoxides / Q000494:pharmacology; D013548:Sweden; D030014:Tabernaemontana / Q000737:chemistry; D014361:Trypsin Inhibitors; D029649:Violaceae / Q000737:chemistry; D000516:alpha-Amylases / Q000037:antagonists & inhibitors
chemical_list: D000893:Anti-Inflammatory Agents; D001688:Biological Products; D010455:Peptides; D010940:Plant Proteins; D010972:Platelet Activating Factor; D014361:Trypsin Inhibitors; D013481:Superoxides; D003571:Cytochalasin B; D009240:N-Formylmethionine Leucyl-Phenylalanine; D001586:Benzoylarginine Nitroanilide; D004074:Digitoxin; D000516:alpha-Amylases; D019272:Leukocyte Elastase; D004072:Digitonin