摘要
目的:研究迷迭香酸对黄嘌呤氧化酶的抑制作用。方法:将20、40、60μg/ml迷迭香酸或1μg/ml阳性对照别嘌呤醇,分别加入黄嘌呤溶液(测尿酸生成量:1 mmol/L;测超氧离子:50μmol/L)和0.1 U/ml黄嘌呤氧化酶中,用生化仪测定5min尿酸生成量和超氧离子生成(NBT显色法)。在1 ml 2×105/ml HL-60细胞悬液中加入100μl 6 mol/L黄嘌呤、100μl0.1 U/ml黄嘌呤氧化酶、500μg/ml迷迭香酸,分别以AnnexinⅤ-PI双标试剂盒法(以1μg/ml别嘌呤醇为阳性对照)或细胞周期法(以100 U/ml SOD为阳性对照)测定细胞凋亡率。结果:迷迭香酸显著抑制尿酸生成和超氧离子引起的NBT显色,两种方法测得其IC50分别为56μg/ml和21μg/ml;对细胞凋亡的抑制率均在40%以上。结论:迷迭香酸是黄嘌呤氧化酶的竞争性抑制剂。
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; 50μmol/L for determining superoxide anions） in the presence of 20, 40 and 60μg/ml rosmarinic acid or allopurinol as positive control. The formation of uric acid was determined by automatic biochemical analyzer 5 rain after reaction and the production of superoxide anions was measured by Nitro Blue Btetrazolium （NBT） reduction. HL-60 cells （1 ml, 2×10^5/ml） were pretreated with xanthine （100μl, 6 mol/L） and xanthine oxidase （100μl, 0.1 U/ml）, then rosmarinic acid （500μg/ml） or allopurinol （1μg/ml, as positive control） （AnnexinV-PI kit） was added to determine the cell apoptosis rate. HL-60 cells （1 ml, 2 × 10^5/ml） were also pretreated with xanthine （100 μl, 6 mol/L） and xanthine oxidase （100μl, 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 IC50being 56μg/ml and 21μg/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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2006年第2期189-191,共3页
Academic Journal of Second Military Medical University
基金
国家自然科学基金(29632050)~~
