Effects of equol on H2O2-induced oxidative stress in primary chicken intestinal epithelial cells
Effects of equol on H2O2-induced oxidative stress in primary chicken intestinal epithelial cells
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DOI：
10.3382/ps/pew034
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发表时间：
2016-06-01
期刊：
影响因子：
4.4
通讯作者：
Gou, Zhongyong
中科院分区：
文献类型：
--
作者：
Lin, Xiajing;Jiang, Shouqun;Gou, Zhongyong
本试验旨在研究马酚对H2O2诱导的鸡肠上皮细胞氧化应激的抗氧化作用。以龄18天的岭南黄肉鸡胚为材料，在Dulbecco改良Eagle’s培养基/F12中培养IEC。分别用0、10、100、500 nM雌马酚预处理细胞24小时，然后再暴露于300 μ M H2O2中24小时。通过细胞显微照片评估氧化损伤，测量细胞增殖、丙二醛（MDA）含量、细胞总超氧化物歧化酶（T-SOD）活性的抗氧化能力，以及Nrf2、Bcl-2、SOD-1、GSH-Px3、Claudin-1的相对表达。300 μ M H2O2处理对细胞损伤严重，显微镜观察显示正常肠上皮细胞较少。与对照组相比，300 μ M H2O2处理显著降低了活细胞数量，而之前用雌马酚处理没有抵消H2O2的作用（P < 0.05）。与单纯H2O2处理的细胞相比，10、100和500 nM雌马酚预处理能显著提高T-SOD活性（P < 0.05）， 10和100 nM雌马酚预处理能显著提高T-SOD活性（P < 0.05）。在100 nM雌马酚预处理的细胞中，Nrf2转录本的相对丰度较对照组增加（P < 0.05），但Bcl2、GSH-Px3或SOD-1的表达未受影响（P < 0.05）。10和100 nM雌马酚预处理显著提高了Claudin-1的转录丰度（P < 0.05）。马酚通过促进抗氧化基因的表达、增加抗氧化酶的活性和增强抗氧化能力来保护内皮细胞免受氧化损伤；100 nM雌马酚似乎是最有效的浓度。
This experiment investigated the antioxidant effects of equol on oxidative stress induced by H2O2 in chicken intestinal epithelial cells (IEC). IEC, from Lingnan yellow broiler chick embryos at embryonic day 18, were cultured in Dulbecco's modified Eagle's medium/F12. Cells were pretreated with 0, 10, 100, or 500 nM equol for 24 h before exposure to 300 mu M H2O2 during a further 24 h. Oxidative damage was assessed by photomicrographs of cells, measuring cell proliferation, malondialdehyde (MDA) content, and antioxidative capacity from cellular total superoxide dismutase (T-SOD) activity, as well as the relative expressions of Nrf2, Bcl-2, SOD-1, GSH-Px3, Claudin-1. Treatment with 300 mu M H2O2 caused serious damage to cells, with fewer normal intestinal epithelial cells, revealed by photomicroscopy. Treatment with 300 mu M H2O2 significantly decreased live cell numbers compared with controls and prior treatment with equol had no effect in offsetting this action of H2O2 (P > 0.05). Compared with the cells treated just with H2O2, pre-treatment with 10, 100 and 500 nM equol significantly enhanced T-SOD activity (P < 0.05), while 10 and 100 nM equol before H2O2 significantly enhanced T-SOD activity compared with the untreated controls (P < 0.05). In cells pre-treated with 100 nM equol, the relative abundance of Nrf2 transcripts increased from the controls (P < 0.05) but expressions of Bcl2, GSH-Px3, or SOD-1 were unaffected (P > 0.05). Pre-treatment with 10 and 100 nM equol significantly increased the transcript abundance of Claudin-1 (P < 0.05). Equol is shown here to protect IECs from oxidative damage by promoting the expression of antioxidant genes, increasing the activities of antioxidant enzymes, and by enhancing antioxidant capacity; 100 nM equol appeared to be the most effective concentration.