Under the assumption
that the endothelium acts as a sensitive signal transduction interface for hemodynamic and oxidative stress, this study supports previous findings in the literature that smoking in itself has direct microvascular effects, and adds to the current knowledge base by showing that it is possible to positively modify microvascular reactivity in “real-life” through daily consumption of an oral antioxidant such as ascorbate and not exclusively in highly experimental settings with supraphysiological doses. The microvascular effects induced by smoking were mitigated by pre-treatment with ascorbate and the reactivity remained within the same time range as that of untreated subjects before smoking a cigarette. Further studies are required on how CHIR-99021 cost these findings may be applied. The early effects of the intense oxidative stress
induced by smoking on the microcirculation are of particular interest as they are AZD8055 clinical trial possibly reversible in contrast to later consequences with irreversible structural changes in blood vessels. The present study adds to the knowledge that smoking by itself has direct microvascular effects and also that it is possible to affect microvascular reactivity with moderate doses of an oral antioxidant such as ascorbate. Further studies are required how these findings may be applied and their clinical relevance. The authors thank Lu Qing, PhD, for valuable assistance. This work was supported by grants from the Karolinska Institute and the Knut and Alice Wallenberg Foundation. The authors have no conflict of interest to declare. “
“Please Cytidine deaminase cite this paper as: Cocks M, Shepherd SO, Shaw CS, Achten J, Costa ML, Wagenmakers AJM. Immunofluorescence microscopy to assess enzymes controlling nitric oxide availability and microvascular blood flow
in muscle. Microcirculation 19: 642–651, 2012. Objective: The net production of NO by the muscle microvascular endothelium is a key regulator of muscle microvascular blood flow. Here, we describe the development of a method to quantify the protein content and phosphorylation of endothelial NO synthase (eNOS content and eNOS ser1177 phosphorylation) and NAD(P)H oxidase expression. Methods: Human muscle cryosections were stained using antibodies targeting eNOS, p-eNOS ser1177 and NOX2 in combination with markers of the endothelium and the sarcolemma. Quantitation was achieved by analyzing fluorescence intensity within the area stained positive for the microvascular endothelium. Analysis was performed in duplicate and repeated five times to investigate CV. In addition, eight healthy males (age 21 ± 1 year, BMI 24.4 ± 1.0 kg/m2) completed one hour of cycling exercise at ∼65%VO2max.