Biopsy coupled to quantitative immunofluorescence: a new method to study the human vascular endothelium.
Limited availability of endothelial tissue is a major constraint when investigating the cellular mechanisms of endothelial dysfunction in patients with metabolic and cardiovascular diseases. We propose a novel approach that combines collection of 200-1,000 endothelial cells from a superficial forearm vein or the radial artery, with reliable measurements of protein expression by quantitative immunofluorescence analysis. This method was validated against immunoblot analysis in cultured endothelial cells. Levels of vascular endothelial cell activation, oxidative stress, and nitric oxide synthase expression were measured and compared in five patients with severe chronic heart failure and in four healthy age-matched subjects. In summary, vascular endothelial biopsy coupled with measurement of protein expression by quantitative immunofluorescence analysis provides a novel approach to the study of the vascular endothelium in humans.
Colombo
Paolo C
PC
Department of Medicine, Division of Cardiology, Albert Einstein College of Medicine, Bronx, New York 10461, USA. lejemtel@aecom.yu.edu
Ashton
Anthony W
AW
Celaj
Sulejman
S
Talreja
Ashok
A
Banchs
Javier E
JE
Dubois
Nicholas B
NB
Marinaccio
Massimo
M
Malla
Shailesh
S
Lachmann
Justine
J
Ware
J Anthony
JA
Le Jemtel
Thierry H
TH
D000368:Aged; D001158:Arteries; D001706:Biopsy; D002303:Cardiac Output, Low / Q000378:metabolism*; D002478:Cells, Cultured; D002908:Chronic Disease; D051546:Cyclooxygenase 2; D004730:Endothelium, Vascular / Q000378:metabolism* / Q000473:pathology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins; D008875:Middle Aged; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D012016:Reference Values; D015203:Reproducibility of Results; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000378:metabolism; D014680:Veins
D007527:Isoenzymes; D008565:Membrane Proteins; D016328:NF-kappa B; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases