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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 explore the effect of different riboflavin levels on the anti-oxidative function of HepG2 cells. Methods The riboflavin-deficient culture conditions were established by customizing riboflavin-free medium and UV-light to remove riboflavin in fetal bovine serum (FBS). On the basis of this, different concentrations of riboflavin (0.76, 3.76, 6.76, 12.76, 24.76 , 48.76nmol / L) were cultured for 96 h. The cell viability and apoptosis rate were measured at different time points. The content of malondialdehyde (MDA) in the culture supernatant was measured at 72h. The alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, as well as glutathione reductase (GR), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) Internal reduced glutathione (GSH) and oxidized glutathione (GSSG) content. Results With the increase of the concentration of riboflavin, the cell viability increased significantly and the apoptosis rate decreased significantly (P <0.05). The activity of ALT and AST decreased significantly, the content of MDA decreased, the activity of GR increased GSH-Px activity was significantly decreased, GSH content was significantly increased, GSSG content was significantly decreased, GSH / GSSG was significantly increased (P <0.05). Most of the above indicators of riboflavin dose effect inflection point in 12.76nmol / L or so. Conclusion The deficiency of riboflavin can lead to the decrease of anti-oxidative function of HepG2 cells, and the concentration of riboflavin maintaining normal anti-oxidant function should be higher than 12.76 nmol / L.