Antioxidant and Free Radical-Scavenging Activity of Choto-san and Its Related Constituents

• Published in: Biological & Pharmaceutical Bulletin Vol. 27; no. 1; pp. 38 - 46
• Main Authors: Mahakunakorn, Pramote, Tohda, Michihisa, Murakami, Yukihisa, Matsumoto, Kinzo, Watanabe, Hiroshi
• Format: Journal Article
• Language: English; Japanese
• Published: Japan The Pharmaceutical Society of Japan 2004; Pharmaceutical Society of Japan; Japan Science and Technology Agency
• Subjects: Animals; antioxidant; Antioxidants - pharmacology; Biphenyl Compounds; Caffeic Acids - pharmacology; Catechin - pharmacology; Cells, Cultured; Choto-san; Chotoko; Drugs, Chinese Herbal - pharmacology; free radical; Free Radical Scavengers - pharmacology; Hydrogen Peroxide - toxicity; Hydroxyl Radical - metabolism; Iron Chelating Agents - pharmacology; Lipid Peroxidation - drug effects; Male; Mice; Mice, Inbred ICR; NG108-15 cell; Oxidants - metabolism; oxidative stress; Picrates - metabolism; Quercetin - pharmacology; Superoxides - metabolism; Thiobarbituric Acid Reactive Substances - metabolism
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.27.38

The antioxidant properties of Choto-san and its related constituents such as Chotoko and Choto-san without Chotoko, and phenolic compounds contained in Chotoko such as epicatechin, caffeic, acid and quercetin were evaluated. In the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay, the scavenging activity of Chotoko (IC50 14.3 μg/ml) was found to be higher than that of Choto-san (IC50 206.2 μg/ml) and Choto-san without Chotoko (IC50 244.3 μg/ml). Epicatechin (IC50 10.4 μM), caffeic acid (IC50 13.8 μM), and quercetin (IC50 7.1 μM) also revealed scavenging activity against DPPH radicals. Choto-san (IC50 67.7 μg/ml) exhibited stronger inhibitory activity against superoxide anion formation than Choto-san without Chotoko (IC50 92.4 μg/ml) but weaker activity than Chotoko (IC50 18.3 μg/ml). The generation of superoxide anion was also inhibited by epicatechin (IC50 175.2 μM), caffeic acid (IC50 141.7 μM), and quercetin (IC50 18.7 μM). In a hydroxyl radical-scavenging experiment, Choto-san (IC50 2.4 mg/ml), Chotoko (IC50 2.2 mg/ml), Choto-san without Chotoko (IC50 2.8 mg/ml), epicatechin (IC50 3.9 mM), caffeic acid (IC50 3.6 mM), and quercetin (IC50 1.9 mM) exhibited activity. In NG108-15 cells, when added simultaneously with H2O2 (500 μM), Choto-san (250 μg/ml), Chotoko (250 μg/ml), Choto-san without Chotoko (500 μg/ml), epicatechin (200 μM), caffeic acid (200 μM), and quercetin (200 μM) effectively protected cells from oxidative damage. In conclusion, the present results provide evidence that Choto-san acts as an antioxidant and cytoprotective agent against oxidative damage, which is due at least partly to the phenolic compounds contained in Chotoko.