Genistein promotes the metabolic transformation of acetaminophen to glucuronic acid in human L-O2, HepG2 and Hep3b cells via the Nrf2/Keap1 pathway
Genistein promotes the metabolic transformation of acetaminophen to glucuronic acid in human L-O2, HepG2 and Hep3b cells via the Nrf2/Keap1 pathway
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DOI：
10.1039/c6fo00889e
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发表时间：
2016-01-01
期刊：
影响因子：
6.1
通讯作者：
Dai Zi-Ling
中科院分区：
文献类型：
--
作者：
Fan Yuan-Jing;Wei Wei;Dai Zi-Ling
本研究旨在探讨金雀异黄素通过Nrf2/Keap1途径调节UGT活性的作用，并阐明其解毒保肝的作用机制。在L-02、HepG2和Hep3b细胞上进行了监测染料木素对对乙酰氨基酚诱导的细胞损伤的保护作用的实验。四甲基偶氮唑蓝、天门冬氨酸氨基转移酶、丙氨酸氨基转移酶、乳酸脱氢酶、谷胱甘肽和谷胱甘肽的测定结果表明，染料木素对对乙酰氨基酚诱导的细胞损伤有明显的保护作用，且呈剂量依赖关系。用不含染料木素的10 mM对乙酰氨基酚处理对照细胞，比较对乙酰氨基酚和染料木素联合作用对UGT1A1、1A6和1A9、Nrf2和Keap1 mRNAs表达以及Nrf2和Keap1蛋白表达的影响。结果表明，金雀异黄素可使Nrf2基因表达增强，Keap1基因表达水平明显降低，且呈剂量依赖关系。同时，UGT mRNA表达增加，其中UGT1A9的表达水平最高。金雀异黄素作用24小时后，对乙酰氨基酚残留量明显减少，对乙酰氨基酚葡醛酸化反应明显增加，且呈剂量依赖关系。
This study aimed to explore the effects of genistein on regulating the activation of UGTs via the Nrf2/Keap1 pathway and to elucidate the underlying mechanisms of detoxification and hepatic protection. Experiments monitoring genistein-induced protection against acetaminophen-induced cell damage were performed in L-02, HepG2 and Hep3b cells. The results of the MTT, AST, ALT, LDH, GSH and GSSG assays showed that genistein evidently protected the cells from acetaminophen-induced injury in a dose-dependent manner. The control cells were treated with 10 mM acetaminophen without genistein to compare with the effects of the combination of acetaminophen and genistein on the expression of UGT1A1, 1A6 and 1A9, Nrf2 and Keap1 mRNAs, as well as the expression of Nrf2 and Keap1 proteins, which were tested by western blotting. The results showed that the expression of the Nrf2 mRNA and protein increased; in contrast, the expression levels of the Keap1 mRNA and protein were obviously reduced by genistein in a dose-dependent manner. Meanwhile, the expression of the UGT mRNA was increased, and UGT1A9 exhibited the highest expression among the three UGTs. Accordingly, the residual acetaminophen content was obviously reduced and acetaminophen glucuronidation increased after 24 hours of treatment with genistein in a dose-dependent manner.