阿斯巴甜和山梨酸钾协同增强 HepG2 细胞内的线粒体 ROS、p53 激活和细胞凋亡。

Synergistic Effects of The Enhancements to Mitochondrial ROS, p53 Activation and Apoptosis Generated by Aspartame and Potassium Sorbate in HepG2 Cells.

机构信息

Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310035, China.

Wenzhou Entry-exit Inspection and Quarantine Bureau, Wenzhou 325027, China.

出版信息

Molecules. 2019 Jan 28;24(3):457. doi: 10.3390/molecules24030457.

The safety of food additives has been widely concerned. Using single additives in the provisions of scope is safe, but the combination of additives, may induce additive, synergy, antagonism and other joint effects. This study investigated the cytotoxicity of aspartame (AT) together with potassium sorbate (PS). Thiazolyl Blue Tetrazolium Bromide (MTT) assay indicated that AT and PS had IC values of 0.48 g/L and 1.25 g/L at 24 h, respectively. High content analysis (HCA) showed that both AT and PS had a negative effect on mitochondrial membrane potential (MMP), reactive oxygen species (ROS) and DNA damage while the joint group behaved more obviously. The biochemical assays revealed typical cell morphological changes and the activation of cytochrome c and caspase-3 verified apoptosis induced by AT together with PS. With dissipation of MMP and increase of cell membrane permeability (CMP), it indicated AT together with PS-induced apoptosis was mediated by mitochondrial pathway. Meanwhile, p53 were involved in DNA damage, and the ratio of Bax/Bcl-2 was increased. Moreover, excessive ROS induced by AT together with PS is a key initiating factor for apoptosis. All these results proved that p53 was involved in apoptosis via mitochondria-mediated pathway and the process was regulated by ROS.

食品添加剂的安全性一直受到广泛关注。在规定的范围内使用单一添加剂是安全的，但添加剂的组合可能会引起添加剂、协同、拮抗等联合作用。本研究探讨了阿斯巴甜（AT）与山梨酸钾（PS）的细胞毒性。噻唑蓝溴化四唑（MTT）检测结果表明，AT 和 PS 在 24 h 时的 IC 值分别为 0.48 g/L 和 1.25 g/L。高内涵分析（HCA）显示，AT 和 PS 均对线粒体膜电位（MMP）、活性氧（ROS）和 DNA 损伤有负向作用，而联合组表现更为明显。生化检测显示典型的细胞形态变化，细胞色素 c 和 caspase-3 的激活证实了 AT 与 PS 共同诱导的细胞凋亡。随着 MMP 的耗散和细胞膜通透性（CMP）的增加，表明 AT 与 PS 诱导的细胞凋亡是通过线粒体途径介导的。同时，p53 参与了 DNA 损伤，Bax/Bcl-2 的比值增加。此外，AT 与 PS 诱导的过量 ROS 是细胞凋亡的关键起始因子。这些结果均表明，p53 通过 ROS 调控的线粒体途径参与细胞凋亡。