Biopsy coupled to quantitative immunofluorescence: a new method to study the human vascular endothelium
Paolo C Colombo, Anthony W Ashton, Sulejman Celaj, Ashok Talreja, Javier E Banchs, Nicholas B Dubois, Massimo Marinaccio, Shailesh Malla, Justine Lachmann, J Anthony Ware, Thierry H Le Jemtel, Paolo C Colombo, Anthony W Ashton, Sulejman Celaj, Ashok Talreja, Javier E Banchs, Nicholas B Dubois, Massimo Marinaccio, Shailesh Malla, Justine Lachmann, J Anthony Ware, Thierry H Le Jemtel
Abstract
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.
Source: PubMed