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当前位置: 首页 > 文章 > 木犀草素与叶酸对黄曲霉毒素B_1致食管上皮细胞毒性及MTHFR基因高甲基化的影响 食品科学 2019 (23) 142-150
Position: Home > Articles > Protective Effect of Luteolin and Folic Acid on Aflatoxin B_1-Induced Cytotoxicity in Esophageal Epithelial Cells and Their Effect on MTHFR Hypermethylation FOOD SCIENCE 2019 (23) 142-150

木犀草素与叶酸对黄曲霉毒素B_1致食管上皮细胞毒性及MTHFR基因高甲基化的影响

作  者：
付凌萌;吴逸;王菁;魏婕;王少康;孙桂菊
单  位：
东南大学公共卫生学院营养与食品卫生学系
关键词：
木犀草素;叶酸;黄曲霉毒素B1;人正常食管上皮细胞;MTHFR基因;甲基化;表观遗传
摘  要：
目的:研究木犀草素(luteolin,LUT)与叶酸(folic acid,FA)对黄曲霉毒素B_1(aflatoxin B_1,AFB_1)诱导损伤的人正常食管上皮细胞(human normal esophageal epithelial cells,HEEC)MTHFR基因甲基化的影响。方法:不同浓度(0、13、25、50、100、200 μmol/L)AFB_1染毒HEEC 24、48、72 h,CCK-8法检测细胞活力;将HEEC分为空白对照组、AFB_1染毒组(200 μmol/L)、LUT干预组(160 μmol/L)、FA干预组(20、200 μmol/L)以及联合干预组(160 μmol/L LUT+20 μmol/L FA、160 μmol/L LUT+200 μmol/L FA),处理24 h后CCK-8法检测细胞活力,流式细胞术检测细胞周期及凋亡,Western blot检测MTHFR蛋白的表达水平,MassARRAY甲基化检测MTHFR基因启动子区甲基化的情况。结果:不同浓度的AFB_1染毒HEEC 24、48、72 h均可以抑制细胞增殖,而经LUT和FA干预后,与AFB_1染毒组相比,LUT及联合干预组细胞周期阻滞显著减少(P<0.05),细胞抑制率及凋亡率显著降低(P<0.05),MTHFR蛋白表达上调有所改善(P<0.05)。且LUT与FA及二者联合对MTHFR基因启动子区高甲基化水平均有降低作用(P<0.05)。结论:AFB_1对HEEC有毒性作用,表现在增殖抑制、周期阻滞,促进凋亡,上调了MTHFR蛋白的表达,LUT干预可减弱这些损伤,对AFB_1所致毒性起到一定的保护作用。AFB_1提高了MTHFR基因启动子区甲基化水平,导致表观遗传的改变。LUT与FA及联合作用可以降低该甲基化水平,改善该表观遗传的改变。
译  名：
Protective Effect of Luteolin and Folic Acid on Aflatoxin B_1-Induced Cytotoxicity in Esophageal Epithelial Cells and Their Effect on MTHFR Hypermethylation
作  者：
FU Lingmeng;WU Yi;WANG Jing;WEI Jie;WANG Shaokang;SUN Guiju;Department of Nutrition and Food Hygiene, School of Public Health, Southeast University;
关键词：
luteolin;;folic acid;;aflatoxin B1;;human normal esophageal epithelial cells;;MTHFR;;methylation;;epigenetic
摘  要：
Objective: To investigate the protective effect of luteolin(LUT) and folic acid(FA) on aflatoxin B_1(AFB_1)-induced injury of human normal esophageal epithelial cells(HEECs) as well as on the methylation of the MTHFR gene. Methods: HEECs were exposed to different concentrations(0, 13, 25, 50, 100 and 200 μmol/L) of AFB_1 for 24, 48 or 72 h, and cell viability was assessed by cell counting kit-8(CCK-8) assay. Subsequently, HEECs were randomly divided into several groups including negative control, AFB_1 poisoning, LUT intervention(160 μmol/L), FA intervention(20 and 200 μmol/L) and joint intervention(160 μmol/L LUT + 20 μmol/L FA, and 160 μmol/L LUT + 200 μmol/L FA). After 24 h of treatment, cell viability was assessed by CCK-8 assay, cell cycle and apoptosis were analyzed by flow cytometry, the protein expression of MTHFR was detected by Western blot, and the methylation of the MTHFR gene promoter was detected by the MassARRAY method. Results: Cell proliferation was inhibited after being exposed to each AFB_1 concentration. Compared with AFB_1 poisoning group, cell cycle arrest and inhibitory and apoptotic rates were significantly reduced and the up-regulation of MTHFR protein expression was decreased in LUT and joint intervention groups(P < 0.05). Moreover, LUT and/or FA interventions reduced the hypermethylation level of the MTHFR gene promoter(P < 0.05). Conclusion: AFB_1 has toxic effects on HEECs as indicated by the inhibition of proliferation, cell cycle arrest, the promotion of apoptosis, and the up-regulation of MTHFR protein expression. LUT ameliorates these injuries and protects against the toxic effects of AFB_1. Moreover, AFB_1 can increase the methylation level of the MTHFR gene promoter, resulting in epigenetic changes. LUT and/or FA intervention can reduce the methylation level and reverse the epigenetic changes.

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