Inhibition of xathine oxidase by rosmarinic acid
- Author:
Yan-Jun SHANG
1
Author Information
1. Department of Biochemistry and Molecular Biology
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(2):189-191
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To study the inhibitory effect of rosmarinic acid on xanthine oxidase. Methods: Xanthine oxidase (0.1 U/ml) was incubated with xanthine (1 mmol/L for determining formation of uric acid; 50 μmol/L for determining superoxide anions) in the presence of 20, 40 and 60 μg/ml rosmarinic acid or allopurinol as positive control. The formation of uric acid was determined by automatic biochemical analyzer 5 min after reaction and the production of superoxide anions was measured by Nitro Blue Btetrazolium (NBT) reduction. HL-60 cells (1 ml, 2×105/ml) were pretreated with xanthine (100 μl, 6 mol/L) and xanthine oxidase (100 μl, 0.1 U/ml), then rosmarinic acid (500 μg/ml) or allopurinol (1 μg/ ml, as positive control) (Annexin V-PI kit) was added to determine the cell apoptosis rate. HL-60 cells (1 ml, 2×105/ml) were also pretreated with xanthine (100 μl, 6 mol/L) and xanthine oxidase (100 μl, 0.1 U/ml), then rosmarinic acid (500 μg/ml) or SOD (100 U/ml, as positive control) (cell cycle method) was added to determine the cell apoptosis rate. Results: Rosmarinic acid obviously inhibited the production of uric acid and superoxide anion-induced reaction in NBT assay, with their IC50 being 56 μg/ml and 21 μg/ml, respectively. The rates of apoptosis inhibition by rosmarinic acid were both over 40% by Annexin V-PI kit and cell cycle method. Conclusion: Rosmarinic acid is a competitive inhibitor of xanthine oxidase.
