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NO x generation by cultured small intestinal epithelial cells

  • Intestinal Disorders, Inflammatory Bowel Disease, Immunology, and Microbiology
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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 NO x generation was determined. NO synthase activity was determined in sonicated supernatants of IEC-6 cells by [14C] arginine conversion to citrulline. NO x generation in subconfluent cultures was greater than in fully confluent cultures, suggesting contact inhibition. NO x generation by IEC-6 cells was significantly increased by CaI and inhibited byl-NAME andl-NNA. LPS, IL-1β, IL-2, IL-8, IFN-8, TFN-α, EGF, TGF-α, bFGF, and PHA significantly increased NO x 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 NO x 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.

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Authors and Affiliations

  1. Gastrointestinal Unit and Center for the Study of Inflammatory bowel Diseases, Massachusetts General Hospital, Boston, Massachusetts

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

Authors
  1. Axel U. Dignass MD
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  2. Daniel K. Podolsky MD
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  3. Daniel Rachmilewitz MD
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Additional information

Dr. Rachmilewitz is on sabbatical leave from Hadassah University Hospital, Jerusalem, Israel.

This work was supported by a grant from the National Institutes of Health (5P30 DK 43351).

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Dignass, A.U., Podolsky, D.K. & Rachmilewitz, D. NO x generation by cultured small intestinal epithelial cells. Digest Dis Sci 40, 1859–1865 (1995). https://doi.org/10.1007/BF02208647

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  • DOI: https://doi.org/10.1007/BF02208647

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