广东工业大学学报 ›› 2024, Vol. 41 ›› Issue (05): 58-65.doi: 10.12052/gdutxb.230137

• 环境科学与工程 • 上一篇    下一篇

桉树林区水库致黑物鞣花酸对人源细胞的毒理效应研究

蒋然1,4, 李心砚2, 周义鸾2, 郭晋川3, 董延军1, 潘越3, 张心凤1, 曾程5   

  1. 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
  • 收稿日期:2023-09-08 出版日期:2024-09-25 发布日期:2024-10-08
  • 通信作者: 李心砚(1994-),女,副教授,博士,主要研究方向为环境毒理学,E-mail:Li_xinyan224@163.com
  • 作者简介:蒋然(1974-),女,正高级工程师,博士,主要研究方向为供水安全与生态毒理学,E-mail:458750084@qq.com
  • 基金资助:
    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)

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

Jiang Ran1,4, Li Xin-yan2, Zhou Yi-luan2, Guo Jin-chuan3, Dong Yan-jun1, Pan Yue3, Zhang Xin-feng1, Zeng Cheng5   

  1. 1. Key Laboratory of the Pearl River Estuary Regulation and Protection of Ministry of Water Resources, Pearl River Water Research Instituion, Guangzhou 510640, China;
    2. School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou 510006, China;
    3. Guangxi Water Conservancy Science Research Institute, Nanning 530013, China;
    4. Guangdong River, Lake Life & Health Engineering Technology Research Center, Pearl River Water Research Instituion, Guangzhou 510640, China;
    5. The Fifth Affiliated Hospital, Guangzhou Medical University, Guangzhou 510180, China
  • Received:2023-09-08 Online:2024-09-25 Published:2024-10-08

HTML

PDF (PC)

72

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1 μmol/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。

关键词: 桉树林区水库, 鞣花酸, 体外实验, 细胞毒性, 联合暴露

Abstract: 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 1 μmol/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 contaminants in southern rural water sources.

Key words: reservoirs of southern eucalyptus forest area, ellagic acid, in vitro, cytotoxicity, combined exposure

中图分类号: 

  • R994.6
[1] 李一平, 罗凡, 郭晋川, 等. 我国南方桉树(Eucalyptus) 人工林区水库突发性泛黑形成机理初探[J]. 湖泊科学, 2018, 30(1): 15-24.
LI Y P, LUO F, GUO J C, et al. Mechanism of “black-water” occurrence in the reservoirs with Eucalyptus plantation, southern China [J]. Journal of Lake Sciences, 2018, 30(1): 15-24.
[2] 王贺亚. 基于3S技术的广西典型水库集水区桉树人工林及其生态系统服务功能的时空动态格局研究 [D]. 上海: 东华大学, 2016.
[3] 杨钙仁, 于婧睿, 苏晓琳, 等. 桉树人工林黑水发生环境及其对鱼类的影响[J]. 西南农业学报, 2016, 29(2): 445-450.
YANG G R, YU J R, SU X L, et al. Occurrence of black water in cut-over areas of eucalyptus plantation and its effect on fish [J]. Southwest China Journal of Agricultural Sciences, 2016, 29(2): 445-450.
[4] 李一平, 罗凡, 李荣辉, 等. 桉树人工林区水体泛黑机理研究进展[J]. 河海大学学报(自然科学版) , 2019, 47(5) : 393-405.
LI Y P, LUO F, LI R H, et al. Review on the mechanism of blackwater in eucalyptus plantation [J]. Journal of Hohai University (Natural Sciences) , 2019, 47(5) : 393-405.
[5] LIANG W Z, CHOU C T, CHENG J S, et al. The effect of the phenol compound ellagic acid on Ca2+ homeostasis and cytotoxicity in liver cells[J]. European Journal of Pharmacology. 2016, 780: 243-251.
[6] OWCZAREK A, RÓZALSKI M, KRAJEWSKA U, et al. Rare ellagic acid sulphate derivatives from the rhizome of geum rivale l. structure, cytotoxicity, and validated HPLC-PDA assay [J]. Applied Sciences, 2017, 400(7): 1-13.
[7] SAMER H H A, MOTHANNA A Q, MOHAMED E Z, et al. Cytotoxicity and antimicrobial activity studies of an Ellagic Acid-Zinc layered hydroxide intercalation compound [J]. Science of Advanced Materials, 2013(5): 1-10.
[8] CEKER S A G, ANAR M, KIZIL H E. Determination of antigenotoxic, proliferative and cytotoxic properties of ellagic acids [J]. The FEBS Journal, 2016, 283(Suppl.1): 160-161.
[9] 李秀宇, 马芹. 水龙骨中没食子酸与鞣花酸的含量测定[J]. 化学研究, 2023, 34(1): 14-19.
LI X Y, MA Q. Determination of gallic acid and ellagic acid in Polypodii Rhizoma [J]. Chemical Research, 2023, 34(1): 14-19.
[10] 周丹水, 张礼行, 廖玮涛, 等. HPLC法测定化香虫茶中没食子酸和鞣花酸的含量[J]. 广东药科大学学报, 2019, 35(3): 373-377.
ZHOU D S, ZHANG L H, LIAO W T, et al. Determination of gallic acid and ellagic acid in Platycarya strobilacea insect tea by HPLC [J]. Journal of Guangdong Pharmaceutical University, 2019, 35(3): 373-377.
[11] 胡贤达, 耿金菊, 于清淼, 等. 使用生物测试方法评价水质毒性的研究进展[J]. 工业水处理, 2021, 41(6): 14-20.
HU X D, GENG J J, YU Q M, et al. Research progress of aquatic toxicity evaluation using bioassays [J]. Industrial Water Treatment, 2021, 41(6): 14-20.
[12] LI N, WANG D H, ZHOU Y Q, et al. Dibutyl phthalate con-tributes to the thyroid receptor antagonistic activity indrinking water processes [J]. Environmental Science & Technology, 2010, 44(17): 6863-6868.
[13] WANG W, WANG M, XU J, et al. Overexpressed GATA3 enhances the sensitivity of colorectal cancer cells to oxaliplatin through regulating MiR-29b [J]. Cancer Cell International, 2020, 20: 1-16.
[14] MENG, D, ZHANG P, ZHANG L, et al. Detection of cellular redox reactions and antioxidant activity assays [J]. Journal of Functional Foods, 2017, 37: 467-479.
[15] LI K, PU K Y, CAI L, et al. Phalloidin-functionalized hyperbranched conjugated polyelectrolyte for filamentous actin imaging in living hela cells [J]. Chemistry of Materials, 2011, 23(8): 2113-2119.
[16] SHEN Y, YAN Y, LU L, et al. Klotho ameliorates hydrogen peroxide-induced oxidative injury in TCMK-1 cells [J]. International Urology and Nephrology, 2018, 50: 787-798.
[17] POLJAK-BLAZI M, JAGANJAC M, SABOL I, et al. Effect of ferric ions on reactive oxygen species formation, cervical cancer cell lines growth and E6/E7 oncogene expression [J]. Toxicology in Vitro, 2011, 25(1): 160-166.
[18] LI W J, JIANG H, SONG N, et al. Dose-and time-dependent α-synuclein aggregation induced by ferric iron in SK-N-SH cells [J]. Neuroscience Bulletin, 2010, 26(3) : 205.
[19] KANDOLA K, BOWMAN A, BIRCH-MACHIN M A. Oxidative stress-a key emerging impact factor in health, ageing, lifestyle and aesthetics[J]. International Journal of Cosmetic Science, 2015, 37 Suppl 2: 1-8.
[20] GUO W, LIU X, LI J, SHEN Y, et al. Prdx1 alleviates cardiomyocyte apoptosis through ROS-activated MAPK pathway during myocardial ischemia/reperfusion injury [J]. International Journal of Biological Macromol, 2018, 112: 608-615.
[21] CHELOMBITKO M A, FEDOROV, A V, ILYINSKAYA, O P, et al. Role of reactive oxygen species in mast cell degranulation [J]. Biochemistry (Mosc) , 2016, 81(12): 1564-157.
[22] EDDAOUDI A, CANNING S L, KATO I. Flow cytometric detection of G0 in live cells by hoechst 33342 and Pyronin Y staining [J]. Methods in Molecular Biology (Clifton, N. J. ), 2018, 1686: 49-57.
[23] KWAN J, WANG H, MUNK S, et al. In high glucose protein kinase C-zeta activation is required for mesangial cell generation of reactive oxygen species [J]. Kidney International, 2005, 68(6): 2526-2541.
[24] CAO M, PENG B, CHEN H, et al. miR-34a induces neutrophil apoptosis by regulating Cdc42-WASP-Arp2/3 pathway-mediated F-actin remodeling and ROS production [J]. Redox Report, 2022, 27(1): 167-175.
[1] 刘迪, 郭清泉, 赵海山, 舒均中, 练英铎, 夏树敏. 丁香酚抗马拉色菌活性、细胞毒性及抑菌机制研究[J]. 广东工业大学学报, 2021, 38(02): 88-93.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2017 《广东工业大学学报》编辑部
地址：广州市东风东路729号广东工业大学 邮编：510090
电话：020-37626653,020-37626139,020-37626396
邮箱：xbzrb@gdut.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发