葛根中黄嘌呤氧化酶抑制剂对GES-1细胞诱导的氧化应激保护作用

Protective Effect of Xanthine Oxidase Inhibitor from Pueraria Lobate Root on Oxidative Stress Induced GES-1 Cells

采用超滤与液相色谱-质谱联用(UF-LC-MS)方法成功地从葛根提取液中筛选并识别出4种黄嘌呤氧化酶(XO)抑制剂,即葛根素、黄豆苷、黄豆苷元和染料木黄酮,其对XO的半数抑制浓度(IC_(50))分别为30.8,5.31,14.5和3.02μg/mL。此外,采用过氧化氢诱导人胃上皮细胞(GES-1)氧化应激模型,研究了XO抑制剂对损伤细胞的保护作用。结果表明,4种XO抑制剂均表现出保护作用,且无细胞毒性。随着4种抑制剂浓度的增加,过氧化氢诱导的细胞内超氧物歧化酶(SOD)活性降低得到恢复,乳酸脱氢酶的释放和细胞内超氧阴离子生成的增加均受到抑制。4种XO抑制剂能显著地提高GES-1细胞的活性,减少氧化应激对GES-1细胞的损伤。

The xanthine oxidase(XO) inhibitor was screened from the extract of Pueraria lobate root by ultrafiltration coupled with liquid chromatography-mass spectrometry(LC-MS).Four XO inhibitors, puerarin, daidzin, daidzein and genistein were identified by this method and half-maximal inhibitory concentration(IC_(50)) values of XO were 30.8, 5.31, 14.5 and 3.02 μg/mL, respectively. The fluorescence spectroscopy was established to study interactions between these compounds and XO. The paper used oxidative stress model of human gastric epithelial cells(GES-1) induced by hydrogen peroxide to study the protective effect of XO inhibitor on injured cells. The results indicated that four XO inhibitors displayed protections and no cytotoxicity.Furthermore, while the concentration of the four inhibitors increased, the decrease of intracellular superoxide dismutase(SOD)activity induced by hydrogen peroxide was restored and the release of lactate dehydrogenase and the increase of intracellular superoxide anion production was inhibited. In conclusion, these four XO inhibitors not only obviously heightened the viability of GES-1 cells but also ameliorated damage of GES-1 cells from oxidative stress.