Development of an in vitro screening assay to test the antiinflammatory properties of dietary supplements and pharmacologic agents

Clin Chem. 2005 Dec;51(12):2252-6. doi: 10.1373/clinchem.2005.056093. Epub 2005 Sep 15.

Abstract

Background: Monocytes and macrophages are critical in atherosclerosis and on stimulation secrete proinflammatory, proatherogenic cytokines such as tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, which have been shown to be present in atherosclerotic lesions. The aim of this study was to develop a rapid in vitro screening assay to test the antiinflammatory effects of different compounds.

Methods and results: THP-1 cells (human monocytic cell line) were stimulated with different concentrations of lipopolysaccharide (LPS; 0 to 1000 microg/L) and for different times (4, 12, and 24 h), and the secretion of proinflammatory cytokines (IL-1, IL-6, and TNF-alpha) was assessed. TNF-alpha secretion was maximum at the lowest LPS concentration (100 microg/L) and at shortest duration of incubation (4 h). Maximum secretion of IL-1beta and IL-6 was achieved at 24 h with higher doses of LPS. Treatment of THP-1 with various test compounds such as dietary supplements (alpha-tocopherol, N-acetylcysteine, catechin and epigallocatechin gallate) as well as pharmacologic agents (statins, peroxisome proliferator-activated receptor-gamma agonists, and an angiotensin II receptor blocker) significantly inhibited LPS-stimulated TNF-alpha release.

Conclusions: The release of TNF-alpha after stimulation of THP-1 cells with LPS is a valid model system to test novel compounds for potential antiinflammatory effects.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Inflammatory Agents / administration & dosage*
  • Anti-Inflammatory Agents / pharmacology*
  • Cell Line, Tumor
  • Dietary Supplements*
  • Dose-Response Relationship, Drug
  • Drug Evaluation, Preclinical / methods*
  • Humans
  • Lipopolysaccharides / pharmacology
  • Tumor Necrosis Factor-alpha / metabolism*

Substances

  • Anti-Inflammatory Agents
  • Lipopolysaccharides
  • Tumor Necrosis Factor-alpha