Measurement of Oxygen Radicals and Lipid Peroxidation in Neural Tissues

Abstract

One of the most completely validated processes involved in secondary tissue damage following acute brain or spinal cord injury and in many chronic neurodegenerative diseases has to do with the pathological formation of reactive oxygen species (ROS) and reactive nitrogen species (RNS). These are generated by multiple mechanisms and give rise to highly reactive oxygen radicals that can damage neuronal, glial, and microvascular elements. Particular interest has centered upon oxygen radical-induced, iron-catalyzed lipid peroxidation (LP) as the principal mechanism of neuronal injury associated with oxygen radicals. Thus, there has been a growing interest in monitoring increased oxygen radical levels as an index of oxidative stress, as well as measuring markers of LP-associated oxidative injury in in vitro and in vivo model systems and neurological patient samples. Accordingly, the purpose of this unit is to provide a variety of methods for the measurement of hydroxyl radical formation and/or LP in nervous tissue or biofluids.

Oxidative stress in ventral prostate, ovary, and breast by 2,4‐dichlorophenoxyacetic acid in pre‐ and postnatal exposed rats

“…Malondialdehyde (MDA) is an end product of the oxidation of polyunsaturated fatty acids, which can react with thiobarbituric acid (TBA) to produce a complex that can be determined spectrophotometrically. Thus, lipid peroxidation was assessed in terms of the thiobarbituric‐acid reactive substances (TBARS) produced, according to Hall et al (2001). Briefly, 1 volume of tissue homogenate and 2 volumes of stock reagent (15% trichloroacetic acid: 0.37% TBA: 0.25 N hydrochloric acid) were thoroughly mixed and heated in boiling water for 15 min.…”

Oxidative stress in rat brain during experimental status epilepticus: effect of antioxidants

“…After centrifugation of homogenates (10,000 g, 15 min, 4 °C) supernatants were used for analysis according to the manufacturer's protocol. Malondialdehyde (MDA) was quantified by a modified thiobarbituric acid (TBA) assay (Hall and Bosken, 2009). Since MDA is chemically unstable, the precursor 1,1,3,3tetramethoxypropane (Merck, Darmstadt, Deutschland), a stable acetal form of MDA, was used to prepare a standard curve; 1 mol of tetramethoxypropane yields a 1 mol of MDA.…”