桉树林区水库致黑物鞣花酸对人源细胞的毒理效应研究  

Research on the Toxic Effects on Human Cells of Natural Melanogenic Substance Ellagic Acid in Reservoirs of Eucalyptus Forest Area

作　　者：蒋然 李心砚 周义鸾 郭晋川 董延军 潘越 张心凤 曾程 Jiang Ran;Li Xin-yan;Zhou Yi-luan;Guo Jin-chuan;Dong Yan-jun;Pan Yue;Zhang Xin-feng;Zeng Cheng(Key Laboratory of the Pearl River Estuary Regulation and Protection of Ministry of Water Resources,Pearl River Water Research Instituion,Guangzhou 510640,China;School of Biomedical and Pharmaceutical Sciences,Guangdong University of Technology,Guangzhou 510006,China;Guangxi Water Conservancy Science Research Institute,Nanning 530013,China;Guangdong River,Lake Life&Health Engineering Technology Research Center,Pearl River Water Research Instituion,Guangzhou 510640,China;The Fifth Affiliated Hospital,Guangzhou Medical University,Guangzhou 510180,China)

机构地区：[1]珠江水利委员会珠江水利科学研究院水利部珠江河口治理与保护重点实验室,广东广州510640 [2]广东工业大学生物医药学院,广东广州510006 [3]广西壮族自治区水利科学研究院,广西南宁530013 [4]珠江水利委员会珠江水利科学研究院、广东省河湖生命健康工程技术研究中心,广东广州510640 [5]广州医科大学第五临床医学院,广东广州510180

出　　处：《广东工业大学学报》2024年第5期58-65,共8页Journal of Guangdong University of Technology

基　　金：广西重点研发计划项目(2020AB22011);广东省重点领域研发计划项目(2020B111153001)。

摘　　要：我国南方桉树林种植区里面的水库在秋冬季存在明显的天然致黑物质,主要为单宁酸、鞣花酸等多羟基酚类化合物及其铁盐络合物,中小型水库承担了农村生活饮用水源的功能,因此有必要评估天然致黑物质的毒性潜在风险。本文模拟南方桉树林区水库水体特有的天然致黑物,以不同浓度的鞣花酸(Ellagic acid,EA)和三价铁(Fe3+)为研究对象,评价暴露于EA和/或Fe3+对人源结直肠腺癌上皮细胞(DLD1)、肝细胞(LO2)和肾上皮细胞(293T)的一般细胞毒性、细胞内活性氧(Reactive Oxygen Species,ROS)水平和细胞形态变化,并确定不同质量浓度的EA和Fe3+的毒性作用特征。结果提示EA和Fe3+在单独暴露和联合暴露条件下表现出不同程度的细胞毒性。联合暴露时,DLD1在较高质量浓度的EA(1 mg/L,10 mg/L)联合1μmol/L Fe3+处理组中细胞活力显著下降,比EA单独暴露时毒性更强。而LO2和293T在联合暴露组中细胞活力均未产生明显变化,但与Fe3+单独暴露相比联合暴露后细胞活力显著下降。氧化应激检测结果发现,与DMSO对照组相比,EA和/或Fe3+暴露均显著增加了ROS水平。其中,DLD1和LO2的ROS结果可能提示EA与Fe3+共同暴露比EA单独暴露时对细胞内ROS生成的影响更大,提高了氧化应激水平。此外,暴露于EA和/或Fe3+对3种细胞的细胞形态没有发现明显变化,但显著抑制了LO2中肌动蛋白F-actin的表达。结果表明,EA和Fe3+的单独或联合暴露对人源细胞具有较强的毒害作用,按影响程度从大到小排序,依次为:肝细胞LO2>肠道细胞DLD1>肾细胞293T。本文为南方农村水源地污染物的精准管控提供了基础依据。Water sources used for rural residential water in reservoirs of the southern Eucalyptus forest area are clearly exposed to natural melanogens such as tannins,ellagic acid and other polyhydroxyphenolic compounds and their iron salt complexes during autumn and winter,making it necessary to evaluate the effect of natural melanogens on human health.In this study,by simulating the natural melanogenic substances from reservoir waters in southern Eucalyptus forests with different concentrations of ellagic acid,trivalent iron were used to evaluate the general cytotoxicity,intracellular reactive oxygen species(ROS)levels,and cellular morphological changes after exposing to ellagic acid(EA)and/or trivalent iron(Fe3+)on colorectal adenocarcinoma epithelial cells(DLD1),hepatocytes(LO2),and human renal epithelial cells(293T).The results showed cytotoxicity in most cases under both individual and combined exposure conditions.Cell viability was significantly reduced in the higher concentration of EA(1 mg/L,10 mg/L)combined with 1μmol/L Fe3+treatment group,which was more toxic than EA exposure alone.The cell viability of LO2 and 293T cells did not change significantly in the combined exposure group,but they were decreased when compared with that of Fe3+exposure alone.The results of oxidative stress assays found that both EA and/or Fe3+exposure significantly increased ROS levels compared with DMSO controls.The ROS results for DLD1,LO2 may suggest that co-exposure had a greater effect on intracellular ROS secretion than when EA was exposed alone.In addition,no significant changes of cell morphology were observed in all treated groups,but the expression of actin F-actin in LO2 was significantly inhibited.The results of this study indicate that ellagic acid and trivalent iron exposure alone or in combination have deleterious effects on human cells,and the toxic effects in order are sorted as follows:hepatocytes(LO2)>intestinal cells(DLD1)>kidney cells(293T).This study provided an important basis for the precise control of contamina

关 键 词：桉树林区水库 鞣花酸 体外实验 细胞毒性 联合暴露 

分 类 号：R994.6[医药卫生—毒理学]