Diclofenac toxicity in human intestine ex vivo is not related to the formation of intestinal metabolites

Xiaoyu Niu, Inge A. M. de Graaf, Miriam Langelaar-Makkinje, Peter Horvatovich, Geny M. M. Groothuis*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

14 Citations (Scopus)

Abstract

The use of diclofenac (DCF), a nonsteroidal anti-inflammatory drug, is associated with a high prevalence of gastrointestinal side effects. In vivo studies in rodents suggested that reactive metabolites of DCF produced by the liver or the intestine might be responsible for this toxicity. In the present study, precision-cut intestinal slices (PCIS) prepared from the jejunum of 18 human donors were used as an ex vivo model to investigate whether DCF intestinal metabolites are responsible for its intestinal toxicity in man. PCIS were incubated with a concentration range of DCF (0-600 A mu M) up to 24 h. DCF (a parts per thousand yen400 A mu M) caused direct toxicity to the intestine as demonstrated by ATP depletion, morphological damage, caspase 3 activation, and lactate dehydrogenase leakage. Three main metabolites produced by PCIS (4'-hydroxy DCF, 5-hydroxy DCF, and DCF acyl glucuronide) were detected by HPLC. Protein adducts were detected by immunohistochemical staining and showed correlation with the intestinal metabolites. DCF induced similar toxicity to each of the samples regardless of the variation in metabolism among them. Less metabolites were produced by slices incubated with 400 A mu M DCF than with 100 A mu M DCF. The addition of the metabolic inhibitors such as ketoconazole, cimetidine, or borneol decreased the metabolite formation but increased the toxicity. The results suggest that DCF can induce intestinal toxicity in human PCIS directly at therapeutically relevant concentrations, independent of the reactive metabolites 4'-OH DCF, 5-OH DCF, or diclofenac acylglucuronide produced by the liver or formed in the intestine.

Original languageEnglish
Pages (from-to)107-119
Number of pages13
JournalArchives of toxicology
Volume89
Issue number1
DOIs
Publication statusPublished - Jan-2015

Keywords

  • Diclofenac
  • Precision-cut intestinal slices
  • Toxicity
  • Metabolism
  • NONSTEROIDAL ANTIINFLAMMATORY DRUGS
  • LIVER PROTEIN ADDUCTS
  • PRECISION-CUT SLICES
  • IN-VITRO
  • ACYL GLUCURONIDE
  • SMALL-BOWEL
  • CYTOCHROME-P450 ENZYMES
  • CAPSULE ENTEROSCOPY
  • INDUCED ENTEROPATHY
  • HUMAN HEPATOCYTES

Cite this