A neutrophil multitarget functional bioassay to detect anti-inflammatory natural products

J Nat Prod. 2002 Jan;65(1):32-41. doi: 10.1021/np010323o.

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.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents / chemistry
  • Anti-Inflammatory Agents / isolation & purification
  • Anti-Inflammatory Agents / pharmacology*
  • Benzoylarginine Nitroanilide / analysis
  • Biological Assay / methods*
  • Biological Products / chemistry
  • Biological Products / isolation & purification
  • Biological Products / pharmacology*
  • Cytochalasin B
  • Digitalis / chemistry
  • Digitonin / chemistry
  • Digitonin / pharmacology
  • Digitoxin / chemistry
  • Digitoxin / isolation & purification
  • Digitoxin / pharmacology
  • Dose-Response Relationship, Immunologic
  • Hemolysis
  • Humans
  • Lamiaceae / chemistry
  • Leukocyte Elastase / analysis*
  • Leukocyte Elastase / antagonists & inhibitors
  • Leukocyte Elastase / metabolism
  • Magnoliopsida / chemistry*
  • N-Formylmethionine Leucyl-Phenylalanine
  • Neutrophil Activation* / drug effects
  • Neutrophils* / drug effects
  • Neutrophils* / enzymology
  • Peptides
  • Plant Proteins / analysis
  • Plant Proteins / chemistry
  • Plant Proteins / pharmacology
  • Plants, Medicinal / chemistry*
  • Plants, Medicinal / classification*
  • Platelet Activating Factor
  • Superoxides / pharmacology
  • Sweden
  • Tabernaemontana / chemistry
  • Trypsin Inhibitors
  • Violaceae / chemistry
  • alpha-Amylases / antagonists & inhibitors

Substances

  • Anti-Inflammatory Agents
  • Biological Products
  • Peptides
  • Plant Proteins
  • Platelet Activating Factor
  • Trypsin Inhibitors
  • Superoxides
  • Cytochalasin B
  • N-Formylmethionine Leucyl-Phenylalanine
  • Benzoylarginine Nitroanilide
  • Digitoxin
  • alpha-Amylases
  • Leukocyte Elastase
  • Digitonin