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目的观察邻苯二甲酸二(2-乙基己)酯(DEHP)刺激人肝癌细胞系HepG2细胞后对细胞糖代谢和脂代谢关键基因表达的影响,探讨DEHP对离体培养HepG2细胞糖脂代谢的毒性。方法取对数生长期HepG2细胞分为DEHP染毒组和对照组,染毒组分别以终浓度为5、10、50、100、500和1 000μmol/L的DEHP染毒,对照组分别予相同终浓度的二甲基亚砜处理。培养24 h后收集细胞,采用实时荧光定量聚合酶链式反应,检测DEHP染毒成功的内源性标志物过氧化物酶体增殖物激活受体α(PPARα),糖代谢关键基因葡萄糖-6-磷酸酶(G-6-Pase)和磷酸烯醇式丙酮酸羧激酶(PEPCK),以及脂代谢关键基因硬脂酰辅酶A去饱和酶1(SCD1)、脂肪酸合成酶、固醇调节原件结合蛋白-1c和乙酰辅酶A羧化酶1的mRNA转录水平。校正检验水准为0.008。结果与自身对照组比较,除了5μmol/L剂量外,其余5个剂量DEHP染毒组HepG2细胞PPARαmRNA转录水平均上调(P<0.008),说明DEHP染毒模型诱导成功。与自身对照组比较,100和500μmol/L DEHP染毒组HepG2细胞的G-6-Pase mRNA转录水平均上调(P≤0.008);6个剂量下,2组HepG2细胞的PEPCK mRNA转录水平分别比较,差异均无统计学意义(P>0.008)。除100μmol/L DEHP染毒组SCD1 mRNA转录水平较对照组下调(P<0.008)外,其余剂量下,2组HepG2细胞的4种脂代谢关键基因的mRNA转录水平分别比较,差异均无统计学意义(P>0.008)。结论 DEHP对糖代谢影响主要体现为促进糖异生限速酶G-6-Pase表达的上调。DEHP对HepG2细胞脂代谢的影响较为有限。
Objective To investigate the effect of DEHP on the expression of key genes of glucose metabolism and lipid metabolism in human hepatocellular carcinoma cell line HepG2 and to investigate the effect of DEHP on the glucose and lipid metabolism of HepG2 cells in vitro Toxicity. Methods HepG2 cells in logarithmic growth phase were divided into DEHP group and control group. The exposed groups were treated with DEHP at final concentrations of 5, 10, 50, 100, 500 and 1 000 μmol / L respectively, and the control groups were given the same Final concentration of DMSO. After cultured for 24 hours, the cells were harvested and the endogenous markers of peroxisome proliferator-activated receptor α (PPARα), the key gene of glucose metabolism, glucose-6 - phosphatase (G-6-Pase) and phosphoenolpyruvate carboxykinase (PEPCK), as well as the key lipid metabolism genes stearoyl-CoA desaturase 1 (SCD1), fatty acid synthase, Protein-1c and acetyl-Coenzyme A carboxylase 1 mRNA transcription levels. The calibration test level is 0.008. Results Compared with the control group, the transcriptional level of PPARα mRNA in HepG2 cells was increased (P <0.008) except for 5μmol / L dose of DEHP, indicating that the model of DEHP was induced successfully. Compared with self-control group, the transcription level of G-6-Pase mRNA in HepG2 cells exposed to 100 and 500 μmol / L DEHP were all increased (P≤0.008). The PEPCK mRNA transcription levels of HepG2 cells in two groups were compared , The difference was not statistically significant (P> 0.008). Except for 100 μmol / L DEHP, the transcriptional level of SCD1 mRNA in HepG2 cells was lower than that in control group (P <0.008). There was no significant difference in mRNA transcription levels of the four key genes of lipid metabolism in HepG2 cells Significance (P> 0.008). Conclusion The effect of DEHP on glycometabolism is mainly due to the up-regulation of G-6-Pase expression. The effect of DEHP on lipid metabolism in HepG2 cells is relatively limited.