Comparison of Mitochondrial Superoxide Detection Ex Vivo/In Vivo by MitoSOX HPLC Method with Classical Assays in Three Different Animal Models of Oxidative Stress

Overview

Journal Antioxidants (Basel)

Date 2019 Oct 31

PMID 31661873

Citations 16

Authors

Sanela Kalinovic

Matthias Oelze

Swenja Kroller-Schon

Sebastian Steven

Ksenija Vujacic-Mirski

Miroslava Kvandova

Isabella Schmal

Ahmad Al Zuabi

Thomas Munzel

Andreas Daiber

Affiliations

Soon will be listed here.

Abstract

Background: Reactive oxygen and nitrogen species (RONS such as HO, nitric oxide) are generated within the organism. Whereas physiological formation rates confer redox regulation of essential cellular functions and provide the basis for adaptive stress responses, their excessive formation contributes to impaired cellular function or even cell death, organ dysfunction and severe disease phenotypes of the entire organism. Therefore, quantification of RONS formation and knowledge of their tissue/cell/compartment-specific distribution is of great biological and clinical importance.

Methods: Here, we used a high-performance/pressure liquid chromatography (HPLC) assay to quantify the superoxide-specific oxidation product of the mitochondria-targeted fluorescence dye triphenylphosphonium-linked hydroethidium (mitoSOX) in biochemical systems and three animal models with established oxidative stress. Type 1 diabetes (single injection of streptozotocin), hypertension (infusion of angiotensin-II for 7 days) and nitrate tolerance (infusion of nitroglycerin for 4 days) was induced in male Wistar rats.

Results: The usefulness of mitoSOX/HPLC for quantification of mitochondrial superoxide was confirmed by xanthine oxidase activity as well as isolated stimulated rat heart mitochondria in the presence or absence of superoxide scavengers. Vascular function was assessed by isometric tension methodology and was impaired in the rat models of oxidative stress. Vascular dysfunction correlated with increased mitoSOX oxidation but also classical RONS detection assays as well as typical markers of oxidative stress.

Conclusion: mitoSOX/HPLC represents a valid method for detection of mitochondrial superoxide formation in tissues of different animal disease models and correlates well with functional parameters and other markers of oxidative stress.

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