Abstract
Changes in the expression of endothelial nitric oxide synthase (eNOS) in the peritoneum could be involved in the peritoneal dysfunction associated with peritoneal inflammation. Demonstrated recently in bovine endothelial cells was the existence of cytosolic proteins that bind to the 3'-untranslated region (3'-UTR) of eNOS mRNA and could be implicated in eNOS mRNA stabilization. The present work demonstrates that eNOS protein is expressed in human endothelial and mesothelial peritoneal cells. Escherichia coli lipopolysaccharide shortened the half-life of eNOS message, reducing eNOS protein expression in peritoneal mesothelial and endothelial cells. Moreover, under basal conditions, human peritoneal samples expressed cytosolic proteins that bind to the 3'-UTR of eNOS mRNA. The cytosolic proteins that directly bind to 3'-UTR were identified as a 60-kD protein. After incubation of human peritoneal samples with lipopolysaccharide, the binding activity of the cytosolic 60-kD protein increased in a time-dependent manner. Studies are now necessary to determine the involvement of this 60-kD protein in the regulation of eNOS expression in peritoneal cells and particularly its involvement in the peritoneal dysfunction associated with inflammatory reactions.
MeSH Terms
3' Untranslated Regions/genetics
Cytosol/metabolism
Escherichia coli
Humans
Immunologic Techniques
Lipopolysaccharides/pharmacology
Nitric Oxide Synthase/genetics,metabolism
Nitric Oxide Synthase Type III
Peritoneum/enzymology
Proteins/metabolism
RNA, Messenger/metabolism
Tumor Necrosis Factor-alpha/physiology
Chemicals
3' Untranslated Regions
Lipopolysaccharides
Proteins
RNA, Messenger
Tumor Necrosis Factor-alpha
NOS3 protein, human
Nitric Oxide Synthase
Nitric Oxide Synthase Type III
Authors & Affiliations
13 authors, click to expand affiliations / ORCID
Arriero María M
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Rodríguez-Feo Juan A
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Celdrán Ángel
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Miguel Lourdes Sánchez DE
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
González-Fernández Fernando
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Fortes José
Department of Pathology, Fundación, Jiménez Díaz, Madrid, Spain.
Reyero Ana
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Frieyro Octavio
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Pinta Juan C DE LA
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Franco Angeles
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Pastor Carlos
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
Casado Santos
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.
López-Farré Antonio
Cardiovascular Research and Hypertension Laboratory, Fundación Jiménez Díaz, Madrid, Spain.