Choi Doo Jin, Kim Sun-Lim, Choi Ji Won, Park Yong Il
Department of Biotechnology, The CUK Agromedical Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Republic of Korea.
National Institute of Crop Science, RDA, Suwon, Gyeonggi-do 441-857, Republic of Korea.
Life Sci. 2014 Jul 25;109(1):57-64. doi: 10.1016/j.lfs.2014.05.020. Epub 2014 Jun 10.
Neuroprotective effects of maysin, which is a flavone glycoside that was isolated from the corn silk (CS, Zea mays L.) of a Korean hybrid corn Kwangpyeongok, against oxidative stress (H2O2)-induced apoptotic cell death of human neuroblastoma SK-N-MC cells were investigated.
Maysin cytotoxicity was determined by measuring cell viability using MTT and lactate dehydrogenase (LDH) assays. Intracellular reactive oxygen species (ROS) were measured using a 2,7-dichlorofluorescein diacetate (DCF-DA) assay. Apoptotic cell death was monitored by annexin V-FITC/PI double staining and by a TUNEL assay. Antioxidant enzyme mRNA levels were determined by real-time PCR. The cleavage of poly (ADP-ribose) polymerase (PARP) was measured by western blotting.
Maysin pretreatment reduced the cytotoxic effect of H2O2 on SK-N-MC cells, as shown by the increase in cell viability and by reduced LDH release. Maysin pretreatment also dose-dependently reduced the intracellular ROS level and inhibited PARP cleavage. In addition, DNA damage and H2O2-induced apoptotic cell death were significantly attenuated by maysin pretreatment. Moreover, maysin pretreatment (5-50 μg/ml) for 2h significantly and dose-dependently increased the mRNA levels of antioxidant enzymes (CAT, GPx-1, SOD-1, SOD-2 and HO-1) in H2O2 (200 μM)-insulted cells.
These results suggest that CS maysin has neuroprotective effects against oxidative stress (H2O2)-induced apoptotic death of human brain SK-N-MC cells through its antioxidative action. This report is the first regarding neuroprotective health benefits of corn silk maysin by its anti-apoptotic action and by triggering the expression of intracellular antioxidant enzyme systems in SK-N-MC cells.
研究从韩国杂交玉米光平玉的玉米须(CS,Zea mays L.)中分离出的黄酮糖苷麦黄酮对氧化应激(H2O2)诱导的人神经母细胞瘤SK-N-MC细胞凋亡性细胞死亡的神经保护作用。
通过MTT和乳酸脱氢酶(LDH)测定法测量细胞活力来确定麦黄酮的细胞毒性。使用二氯荧光素二乙酸酯(DCF-DA)测定法测量细胞内活性氧(ROS)。通过膜联蛋白V-FITC/PI双重染色和TUNEL测定法监测凋亡性细胞死亡。通过实时PCR测定抗氧化酶mRNA水平。通过蛋白质印迹法测量聚(ADP-核糖)聚合酶(PARP)的裂解。
麦黄酮预处理降低了H2O2对SK-N-MC细胞的细胞毒性作用,表现为细胞活力增加和LDH释放减少。麦黄酮预处理还剂量依赖性地降低了细胞内ROS水平并抑制了PARP裂解。此外,麦黄酮预处理显著减轻了DNA损伤和H2O2诱导的凋亡性细胞死亡。此外,麦黄酮预处理(5-50μg/ml)2小时显著且剂量依赖性地增加了H2O2(200μM)损伤细胞中抗氧化酶(CAT、GPx-1、SOD-1、SOD-2和HO-1)的mRNA水平。
这些结果表明,CS麦黄酮通过其抗氧化作用对氧化应激(H2O2)诱导的人脑SK-N-MC细胞凋亡性死亡具有神经保护作用。本报告首次报道了玉米须麦黄酮通过其抗凋亡作用和触发SK-N-MC细胞内抗氧化酶系统的表达而具有的神经保护健康益处。