核黄素对HepG2细胞抗氧化功能的影响  

EFFECTS OF RIBOFLAVIN ON ANTIOXIDANT FUNCTION IN HEPG2 CELLS

作　　者：辛中豪 高蔚娜[2] 蒲玲玲[2] 姚站馨[2] 王亚雯[1,2] 郭长江[1,2] 

机构地区：[1]广西医科大学公共卫生学院,南宁530021 [2]军事医学科学院卫生学环境医学研究所,天津300050

出　　处：《营养学报》2016年第4期381-385,共5页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.81470147)

摘　　要：目的探索不同核黄素水平对HepG2细胞抗氧化功能的影响。方法通过定制无核黄素培养基、紫外线清除胎牛血清(FBS)中核黄素建立核黄素缺乏培养条件,在此基础上,以不同浓度核黄素(0.76、3.76、6.76、12.76、24.76、48.76nmol/L)培养HepG2细胞,共培养96 h,于不同时间点取样测定细胞活力、细胞凋亡率,于72h测定培养上清液的丙二醛(MDA)含量、丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)活性,以及细胞中谷胱甘肽还原酶(GR)、谷胱甘肽过氧化物酶(GSH-Px)与超氧化物歧化酶(SOD)活性,细胞内还原型谷胱甘肽(GSH)与氧化型谷胱甘肽(GSSG)含量。结果随着核黄素浓度增加,细胞活力显著上升、细胞凋亡率显著下降(P<0.05),培养液中ALT、AST活性显著下降、MDA含量显著下降,细胞内SOD活性显著上升、GR活性显著上升、GSH-Px活性显著下降、GSH含量显著增加、GSSG含量显著减少、GSH/GSSG显著升高(P<0.05)。上述大部分指标的核黄素剂量效应拐点在12.76nmol/L左右。结论核黄素不足可导致HepG2细胞的抗氧化功能下降,维持正常抗氧化功能的核黄素浓度应高于12.76nmol/L。Objective To study the effects of riboflavin on antioxidant function in HepG2 cells. Methods A riboflavin deficient HepG2 cell model was established by using ultraviolet radiation to destroy riboflavin in fetal bovine serum and a riboflavin free medium. The cells were then cultured in the media containing 0.76, 3.76, 6.76, 12.76, 24.76, or 48.76 nmol/L of riboflavin. The changes of cell viability and apoptosis rate were assessed at different time points during 96h culture. In the meantime, extracellular alanine aminotransferase（ALT）, aspartate aminotransferase（AST）, malonaldehyde（MDA）, glutathione （GSH）, oxidized glutathione （GSSG） and intracellular glutathione reductase（GR）, superoxide dismutase（SOD）, glutathione peroxidase（GSH-Px） were assayed biochemically at 72h. Results Compared with the cells cultured in the medium containing 0.76nmol/L riboflavin, the cell viability was significantly increased and apoptosis rate significantly decreased in cells cultured in the media containing more than 12.76nmol/L riboflavin from 48h to 72h （P〈0.05）. Meanwhile, significantly decreased activities of ALT, AST, GSH-Px, contents of MDA, GSSG, as well as significantly increased activities of SOD, GR, content of GSH and ratio of GSH to GSSG were observed in the cells cultured in the media containing more than 12.76nmol/L riboflavin （P〈0.05）. Breaking point calculation showed that the riboflavin showed a significant effect when its concentration was close to 12.76nmol/L. Conclusion Riboflavin is vital in maintaining antioxidant function in HepG2 cells and the concentration of riboflavin that can maintain normal antioxidant function should be more than 12.76nmol/L.

关 键 词：核黄素 HEPG2细胞 抗氧化 

分 类 号：R151[医药卫生—营养与食品卫生学]