Effect of antioxidants on chemiluminescence produced by reactive oxygen species

Abstract

Luminol chemiluminescence was used to evaluate the scavenging of superoxide, hydroxyl and alkoxy radicals by four antioxidants: dipyridamole, diethyldithiocarbamic acid, (+)catechin, and ascorbic acid. Different concentrations of these compounds were compared with well-known oxygen radical scavengers in their capacity to inhibit the chemiluminescence produced in the reaction between luminol and specific oxygen radicals. Hydroxyl radicals were generated using the Fenton reaction and these produced chemiluminescence which was inhibited by diethyldithiocarbamate. Alkoxy radicals were generated using the reaction of tert-butyl hydroperoxide and ferrous ion and produced chemiluminescence which was inhibited equally by all of the compounds tested. For the determination of superoxide scavengers we describe a new, simple, economic, and rapid chemiluminescence method consisting of the reaction between luminol and horseradish peroxidase (HRP). With this method it was found that 40 nmol/l dipyridamole, 0.18 mumol/l ascorbic acid, 0.23 mumol/l (+)catechin, and 3 mumol/l diethyldithiocarbamic acid are equivalent to 3.9 ng/ml superoxide dismutase (specific scavenger of superoxide) in causing the same degree of chemiluminescence inhibition. These results not only indicated that the antioxidative properties of these compounds showed different degrees of effectiveness against a particular radical but also that they may exert their action against more than one radical.

Global Cerebral Ischemia in the Rat: Online Monitoring of Oxygen Free Radical Production Using Chemiluminescence in vivo

“…, 1991)]. It has been used to measure ROS production in cell-free systems (Pascual and Romay, 1992), single cells (Suematsu et ai. , 1989), cultured cells (Caraceni et ai.…”

Antioxidant and anti-inflammatory properties of C-phycocyanin from blue-green algae

“…The method used was described by Pascual and Romay [9]. In brief, to 800 L of a mixture containing: 50 mM K 2 HPO 4 -KH 2 PO 4 buffer pH 7.8, 2 mM EDTA and 0.1 mM luminol, 0.1 mL of distilled water, sample or specific scavenger (DMSO) was added.…”

“…Determination of alkoxyl radical scavenging activity was performed by measuring the inhibition of the CL produced by the reaction of t-BOOH with ferrous ions in the presence of luminol, as previously described [9]. The reaction mixture consisted of: 800 L of 50 mM glycine buffer pH 8.6, 50 mM SDS, 0.025 mM luminol.…”

Protective effects of C-phycocyanin on alcohol-induced acute liver injury in mice

“…TG cannot be removed thereby aggravating liver cell damage, causing abnormal liver function, hepatic tissue fat metabolism, extensive vacuolar degeneration and necrosis of liver cells, which fi nally leads to extensive swelling and cell death. Therefore, the key to prevent ethanol liver injury occurrence and development is to remove the free radicals, inhibit lipid peroxidation, protect liver cells and restore normal liver function (Pascual and Romay, 1992).…”