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Curcumin Effect on Copper Transport in HepG2 Cells

机译:姜黄素对HepG2细胞铜转运的影响

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摘要

Background and Objective: In Wilson’s disease, copper metabolism is impaired due to defective copper transporting protein ATP7B, resulting in copper accumulation in liver and brain and causing damage to liver and brain tissues. Published data suggest that one of the possible treatments for Wilson’s disease is curcumin—a compound found in the root of Curcuma longa. In this study, we tested whether curcumin affects copper transport and excretion in HepG2 hepatocytes carrying wildtype ATP7B. Materials and Methods: We examined the impact of 5 µM and 25 µM curcumin on the transport of copper in HepG2 cells incubated with 20 µM and 100 µM CuCl2, as well as copper excretion from cells. First, immunofluorescent staining and co-localization analysis were carried out in HepG2 cells using confocal laser scanning microscope and Nikon NIS Elements software. Second, a concentration of copper extracted into cell culture medium was determined using atomic absorption spectrometry. Results: The analysis of the co-localization between Golgi complex and ATP7B revealed that both 5 µM and 25 µM doses of curcumin improve the ability of liver cells to transport copper to plasma membrane at 20 µM CuCl2, but not at 100 µM CuCl2 concentration. However, atomic absorption spectrometry showed that curcumin rather promotes copper absorption into liver cell line HepG2 than excretion of it. Conclusions: Curcumin accelerates the transport of copper within liver cells, but does not promote copper excretion from HepG2 cells.
机译:背景与目的:在威尔逊氏病中,由于铜转运蛋白ATP7B缺陷导致铜代谢受损,导致铜在肝和脑中蓄积,并损害肝和脑组织。公开数据表明,针对威尔逊氏病的一种可能的治疗方法是姜黄素,姜黄素是一种在姜黄根中发现的化合物。在这项研究中,我们测试了姜黄素是否影响携带野生型ATP7B的HepG2肝细胞中的铜转运和排泄。材料和方法:我们研究了5 µM和25 µM姜黄素对用20 µM和100 µM CuCl2培养的HepG2细胞中铜转运以及细胞铜排泄的影响。首先,使用共聚焦激光扫描显微镜和尼康NIS Elements软件在HepG2细胞中进行免疫荧光染色和共定位分析。其次,使用原子吸收光谱法确定提取到细胞培养基中的铜的浓度。结果:对高尔基复合体和ATP7B之间共定位的分析表明,在20 µM CuCl2浓度下,5 µM和25 µM剂量的姜黄素均可提高肝细胞将铜转运至质膜的能力,而在100 µM CuCl2浓度下则不会。然而,原子吸收光谱法显示姜黄素比铜的排泄更能促进铜吸收到肝细胞系HepG2中。结论:姜黄素促进肝细胞内铜的转运,但不促进HepG2细胞的铜排泄。

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