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Nitric oxide inhibition of chronic Hcy‐induced endothelial cell (EC) activation is cGMP‐independent
Author(s) -
Graham Anne Margaret,
Alkhoury Kenan,
Naseem Khalid,
Parkin Susan Mary
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.568.1
Subject(s) - nitric oxide , chemistry , cell adhesion molecule , umbilical vein , hyperhomocysteinemia , immunofluorescence , microbiology and biotechnology , cell adhesion , endothelium , endothelial stem cell , cell , in vitro , homocysteine , immunology , medicine , antibody , biochemistry , biology , organic chemistry
Hyperhomocysteinemia (hHcy) upregulates expression of cell adhesion molecules (CAM) on EC which recruits leukocytes and initiates atherosclerotic lesion development. Nitric oxide (NO) is anti‐adhesive for leukocytes. Objective to study effects of exogenous NO on hHcy CAM expression in human umbilical vein endothelial cells (HUVEC) and recruitment of leukocytes under flow. HUVEC were cultured for 5‐9 days in medium containing 1mM Hcy or Cysteine (negative control). Some Hcy‐stimulated cells were treated with NO donor (S‐nitrosoglutathione: GSNO: 20µM, 10min) in absence of presence of a cGMP inhibitor (ODQ 40µM, 30min). Confluent cells were subjected to flowing neutrophils (1x10 6 cells/ml) at 1.1dynes. Tethered, rolled, fixed and transmigrated neutrophils on HUVEC were counted. Immunofluorescence antibodies against ICAM‐1, E‐selectin and P‐selectin quantified adhesion molecule expression. Hcy induced expression of ICAM‐1, E‐selectin and P‐selectin (p<0.05) and significantly increased interactions between neutrophils and HUVEC while cysteine had no effect. When Hcy‐treated HUVEC were exposed to GSNO with or without ODQ, numbers of all interactions decreased dramatically but significantly decreased only P‐selectin expression (p<0.05). NO inhibits HUVEC: leukocyte interaction through inhibiting the expression of P‐selectin in a cGMP‐independent manner. Funding: HRUK, ORSAS