Increased free radical production in hypertension due to increased expression of the NADPH oxidase subunit p22(phox) in lymphoblast cell lines

Journal of Hypertension
Andrew I PettitLeong L Ng

Abstract

To confirm increased production of reactive oxygen species (ROS) in hypertension, to demonstrate the source of ROS and to analyse NADPH oxidase subcomponent expression in hypertension. A lymphoblast model was used, as this has previously been used in the study of hypertension and of NADPH oxidase. Chemiluminescence (CL) was chosen to assay ROS production, as it is simple and sensitive. Lymphocytes from 12 hypertensive patients (HT), and 12 age- and sex-matched normotensive (NT) subjects, were immortalized. Luminol, isoluminol and Cypridina luciferin analogue (CLA) CL were used to assay ROS production. NADPH oxidase subunits were measured by Western blot analysis. Stimulation with 50 micromol/l arachidonic acid (AA) resulted in increased ROS production in HT cell lines with luminol, CLA and isoluminol CL. Stimulation with 500 nmol/l 12-O-tetradecanoylphorbol-13-acetate (TPA) produced a detectable increase in HT ROS production with luminol and with CLA, whereas there was no significant difference with isoluminol. The ROS production was abolished by diphenyleneiodonium chloride (DPI) but not by rotenone, indicating that a non-mitochondrial flavoprotein such as NADPH oxidase is the source of ROS. Analysis of NADPH oxidase subcompon...Continue Reading

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Citations

Jul 27, 2006·Amino Acids·C Cortés-RojoA Saavedra-Molina
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