林丹暴露诱导肝脏HepG2细胞的毒性效应研究

doi:10.7523/j.issn.2095-6134.2021.06.005

中国科学院大学学报 ›› 2021, Vol. 38 ›› Issue (6): 750-757.DOI: 10.7523/j.issn.2095-6134.2021.06.005

林丹暴露诱导肝脏HepG2细胞的毒性效应研究

段珊珊^1,3, 王苑郦², 杨建正^1,3, 丁文军², 贺军辉¹

1. 中国科学院理化技术研究所功能纳米材料实验室微纳材料与技术研究中心, 北京 100190;
2. 中国科学院大学生命科学学院环境与健康实验室, 北京 100049;
3. 中国科学院大学, 北京 100049

收稿日期:2021-02-10 修回日期:2021-04-26 发布日期:2021-11-16
通讯作者: 丁文军, 贺军辉
基金资助:
国家重点研发计划项目（2017YFA0207102）、国家自然科学基金重大研究计划培育项目（91963104）和国家重点研发计划前沿科技创新专项（2019QY（Y）0503）资助

Cytotoxicity of lindane in hepatoma HepG2 cells

DUAN Shanshan^1,3, WANG Yuanli², YANG Jianzheng^1,3, DING Wenjun², HE Junhui¹

1. Functional Nanomaterials Laboratory, Center for Micro/nanomaterials and Technology, Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Beijing 100190, China;
2. Laboratory of Environment and Health, College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China;
3. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2021-02-10 Revised:2021-04-26 Published:2021-11-16

摘要/Abstract

摘要： 有机氯农药林丹具有高毒性、亲脂性、化学稳定性和生物富集性，曾被广泛应用于农业生产中。林丹严重威胁着生态环境和生物安全，体内富集后造成肝功能损伤，但其对肝脏的毒性效应尚不十分清楚。在本研究中，肝脏HepG2细胞经不同浓度林丹暴露处理24 h后，使用噻唑蓝（MTT）、2，7-二氯荧光素（DCF）和免疫荧光方法分别检测细胞活力、细胞内活性氧（ROS）和自噬生成；并用实时定量PCR和Western blot分别检测细胞IL-6、IL-8、IL-1β、TNF-α、NF-κB、Beclin1、ULK1 mRNA和P-p65、IL-1β、P62、LC3蛋白质的表达水平。结果表明，高浓度林丹暴露显著抑制HepG2细胞活力，升高细胞内的ROS和乳酸脱氢酶（LDH）生成水平，降低过氧化氢酶（CAT）活性，并明显上调TNF-α、IL-6、IL-1β、IL-8、NF-κB mRNA和P-p65、IL-1β、P62、LC3、NF-κB蛋白质表达水平。以上结果表明，林丹诱导肝脏HepG2细胞的氧化应激和自噬，并激活NF-κB信号通路引起细胞的炎症反应。

关键词: 林丹, HepG2细胞, 氧化应激, 炎症反应, 自噬

Abstract: Lindane, as one of persistent organic pollutants (POPs), has high toxicity, lipophilicity, chemical stability, and bioaccumulation. The use of lindane seriously threatens ecological environment and biosafety in China. Lindane bioaccumulation can cause liver function damage, but its toxic effects on liver have not been fully understood. In this study, we have investigated effects of lindane-induced oxidative stress on inflammation and autophagy in HepG2 cells. HepG2 cells were exposed to lindane for 24 h. Cell viability, intracellular reactive oxygen species (ROS), and autophagy were determined by thiazolyl blue (MTT) assay, 2,7-dichlorofluorescein (DCF) and immunofluorescence, respectively. The expression levels of IL-6, IL-8, IL-1β, TNF-α, NF-κB, Beclin1, ULK1 mRNA and P-P65, IL-1β, P62, LC3 protein were detected by real-time quantitative PCR and Western blot, respectively. We found that high-concentration lindane significantly decreased the cell viability, triggered an increase in both intracellular ROS and lactate dehydrogenase (LDH) levels, and decreased the catalase (CAT) activity. Moreover, the expression levels of TNF-α, IL-6, IL-1β, IL-8, NF-κB mRNA and P-p65, IL-1β, P62, LC3, and NF-κB protein were significantly up-regulated after lindane exposure. These results indicated that the lindane exposure causes oxidative stress and autophagy as well as inflammatory through activating NF-κB signaling pathway in HepG2 cells.

Key words: lindane, HepG2 cell, oxidative stress, inflammation, autophagy

中图分类号:

D610.10

段珊珊, 王苑郦, 杨建正, 丁文军, 贺军辉. 林丹暴露诱导肝脏HepG2细胞的毒性效应研究[J]. 中国科学院大学学报, 2021, 38(6): 750-757.

DUAN Shanshan, WANG Yuanli, YANG Jianzheng, DING Wenjun, HE Junhui. Cytotoxicity of lindane in hepatoma HepG2 cells[J]. Journal of University of Chinese Academy of Sciences, 2021, 38(6): 750-757.

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林丹暴露诱导肝脏HepG2细胞的毒性效应研究

Cytotoxicity of lindane in hepatoma HepG2 cells

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