Department of Public Health, College of Medicine, Shihezi University, Shihezi 832000, Xinjiang, China.
Biomed Res Int. 2019 Jul 4;2019:3924581. doi: 10.1155/2019/3924581. eCollection 2019.
This study investigated the effects of proanthocyanidins (PC) on arsenic methylation metabolism and efflux in human hepatocytes (L-02), as well as the relationships between PC and GSH, MRP1 and other molecules. Cells were randomly divided into blank control group, arsenic trioxide exposure group (ATO, AsO, 25mol/L), and PC-treated arsenic exposure group (10, 25, 50mg/L). After 24/48h, the contents of different forms of arsenic were determined, and the methylation indexes were calculated. Intracellular S-adenosyl methionine (SAM), arsenic (+3 oxidation state) methyltransferase (AS3MT), multidrug resistance-associated protein 1 (MRP1), and reduced glutathione (GSH) were ascertained. Changing trends were observed and the correlation between arsenic metabolism and efflux related factors and arsenic metabolites was analyzed. We observed that cells showed increased levels of content/constituent ratio of methyl arsenic, primary/secondary methylation index, methylation growth efficiency/rate, and the difference of methyl arsenic content in cells and culture medium (P<0.05, resp.). Compared with ATO exposure group, the intracellular SAM content in PC-treated group decreased, and the contents of GSH, AS3MT, and MRP1 increased (P<0.05, resp.). There was a positive correlation between the content of intracellular GSH/AS3MT and methyl arsenic. The content of MRP1 was positively correlated with the difference of methyl arsenic content in cell and culture medium; conversely, the SAM content was negatively correlated with intracellular methyl arsenic content (P<0.05, resp.). Taken together, these results prove that PC can promote arsenic methylation metabolism and efflux in L-02 cells, which may be related to the upregulation of GSH, MRP1, and AS3MT levels by PC.
本研究旨在探讨原花青素（PC）对人肝细胞（L-02）砷甲基化代谢和外排的影响，以及 PC 与 GSH、MRP1 等分子之间的关系。细胞随机分为空白对照组、三氧化二砷暴露组（ATO，AsO，25μmol/L）和 PC 处理的砷暴露组（10、25、50mg/L）。24/48h 后，测定不同形态砷的含量，计算甲基化指标。测定细胞内 S-腺苷甲硫氨酸（SAM）、砷（+3 氧化态）甲基转移酶（AS3MT）、多药耐药相关蛋白 1（MRP1）和还原型谷胱甘肽（GSH）的含量。观察变化趋势，分析砷代谢与外排相关因素及砷代谢物之间的相关性。结果显示，细胞内甲基砷的含量/组成比、初级/次级甲基化指数、甲基化生长效率/速率以及细胞内外甲基砷含量的差异均增加（P<0.05）。与 ATO 暴露组相比，PC 处理组细胞内 SAM 含量降低，GSH、AS3MT 和 MRP1 含量增加（P<0.05）。细胞内 GSH/AS3MT 含量与甲基砷含量呈正相关，MRP1 含量与细胞内外甲基砷含量差异呈正相关，SAM 含量与细胞内甲基砷含量呈负相关（P<0.05）。综上所述，PC 可促进 L-02 细胞砷的甲基化代谢和外排，这可能与 PC 上调 GSH、MRP1 和 AS3MT 水平有关。
