NOx generation by cultured small intestinal epithelial cells

Axel U. Dignass, Daniel K. Podolsky, Daniel Rachmilewitz

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

The effect of cytokines, growth factors, mitogens, and bacterial products on nitric oxide (NO) generation by monolayers of small intestinal epithelial cells-6 (IEC-6) cells was evaluated. Subconfluent IEC-6 cells were maintained in DMEM containing 5% fetal calf serum and after 16-24 hr of incubation, the medium was replaced with fresh medium in the presence or absence of calcium ionophore (CaI), l-NAME, l-NNA, individual growth factors, cytokines, or mitogens. After 72 hr of culture, the media supernatant was collected and NOx generation was determined. NO synthase activity was determined in sonicated supernatants of IEC-6 cells by [14C] arginine conversion to citrulline. NOx generation in subconfluent cultures was greater than in fully confluent cultures, suggesting contact inhibition. NOx generation by IEC-6 cells was significantly increased by CaI and inhibited by l-NAME and l-NNA. LPS, IL-1β, IL-2, IL-8, IFN-8, TFN-α, EGF, TGF-α, bFGF, and PHA significantly increased NOx generation. NO synthase activity in IEC-6 cells (4.2±1.7 pmol/min/106 cells) was NADPH dependent. These results suggest that stimulation of NOx generation by intestinal epithelial cells through cytokine bacterial products and mitogens may be one of the mechanisms responsible for their effects in the intestinal tract.

Original languageEnglish (US)
Pages (from-to)1859-1865
Number of pages7
JournalDigestive Diseases and Sciences
Volume40
Issue number9
DOIs
StatePublished - Sep 1 1995

Keywords

  • epithelial cells
  • nitric oxide
  • small intestine

ASJC Scopus subject areas

  • Physiology
  • Gastroenterology

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