Benzo[k] fluoranthene induces male germ cell damage

Dandan WANG; Furong WANG; Xiaoyan YANG; Niya ZHOU; Huifang YANG; Jia CAO

doi:10.16016/j.2097-0927.202111178

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Benzo[k] fluoranthene induces male germ cell damage

Basic Medicine | 更新时间：2026-01-26

- Benzo[k] fluoranthene induces male germ cell damage
- Journal of Army Medical University Vol. 44, Issue 8, Pages: 740-748(2022)
- 作者机构：
  
  1.750000 银川，宁夏医科大学公共卫生与管理学院卫生毒理学教研室
  2.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系毒理学研究所
  3.400021 重庆，重庆市妇幼保健院
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81630087);the Joint Project of Medical Scientific Research of Chongqing Science and Technology Commission and Chongqing Health Commission(2021MSXM123)
- DOI：10.16016/j.2097-0927.202111178
  CLC： R-33;R321.1;R994.6
- Received：24 November 2021，
  
  Revised：2022-01-27，
  
  Published：30 April 2022
- 稿件说明：
移动端阅览
Dandan WANG, Furong WANG, Xiaoyan YANG, et al. Benzo[k] fluoranthene induces male germ cell damage[J]. Journal of Army Medical University, 2022, 44(8): 740-748.
DOI：

Dandan WANG, Furong WANG, Xiaoyan YANG, et al. Benzo[k] fluoranthene induces male germ cell damage[J]. Journal of Army Medical University, 2022, 44(8): 740-748. DOI： 10.16016/j.2097-0927.202111178.

摘要

目的

建立高环多环芳烃的代表性化合物苯并[k]荧蒽(BkF)致小鼠精母细胞(GC-2spd)损伤模型，初步探讨BkF引起GC-2细胞的损伤效应及机制。

方法

通过比较毒理基因组学数据库(Comparative Toxicogenomics Database

CTD)筛选与BkF相关的男性生殖系统疾病的基因，对其进行功能和通路富集，探索可能的通路机制。以终浓度为0、10、20、40、80 μmol/L BkF处理GC-2细胞，检测细胞周期、氧化应激水平及DNA损伤效应，采用Western blot检测关键基因及相关通路核心分子的蛋白表达水平。

结果

CTD数据库中筛选出与BkF相关生殖系统疾病的关键基因为CYP1A1和CYP17A1。GO富集分析发现其在男性生殖系统、细胞周期阻滞、氧化应激等生物过程中明显富集，KEGG富集分析发现其在细胞色素P450通路中富集明显(

0.01)。BkF染毒GC-2细胞72 h后，细胞增殖活力较对照组明显下降，且存在剂量-效应关系。染毒剂量高于40 μmol/L后细胞周期阻滞明显(

0.05)，且染毒组细胞内ROS水平(

0.05)、DNA损伤标志物γ-H2AX蛋白表达量(

0.01)呈剂量依赖性增加。与对照组相比，染毒组细胞CYP1A1和CYP17A1的蛋白表达显著增加(

0.05)。加入AhR抑制剂CH223191后，染毒组CYP1A1和CYP17A1的表达明显下调(

0.05)。

结论

BkF通过诱导AhR受体激活，引起下游基因CYP1A1和CYP17A1的表达升高、细胞内ROS增加，从而导致GC-2细胞氧化应激，造成遗传损伤。

Abstract

Objective

To establish a damage model of mouse spermatocyte GC-2spd cells induced by benzo[k] fluoranthene (BkF)

a representative compound of high-molecular weight polycyclic aromatic hydrocarbons

so as to investigate the effect and mechanism of BkF on GC-2 cells.

Methods

Related genes of male reproductive diseases associated with BkF were screened in Comparative Toxicogenomics Database (CTD)

and functional and pathway enrichment analysis is conducted subsequently to explore the possible pathway mechanism. Moreover

GC-2 cells were treated with BkF at a final concentration of 0

40 or 80 μmol/L for 72 h

and then the cell cycle

oxidative stress and DNA damage effect were detected. Western blotting was used to determine the protein levels of the key genes as well as the core molecules of related pathways.

Results

The key genes CYP

1A1 and CYP17A1 associated with BkF-related reproductive system diseases were screened out from the CTD database. Gene Ontology (GO) enrichment analysis found that they were involved in male reproductive system and cell cycle arrest

obviously enriched in biological processes such as oxidative stress. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that these genes were significantly enriched in cytochrome P450 pathway (

0.01). After GC-2 cells were exposed to BkF for 72 h

the proliferation was significantly decreased in a dose-dependent manner

as compared with the control group. When the exposure dose was higher than 40 μmol/L

the cell cycle arrest was obvious (

0.05); Both the intracellular reactive oxygen species (ROS) level (

0.05) and the protein level of DNA damage marker γ-H2AX (

0.01) were remarkably increased in a dose-dependent manner. In addition

the protein levels of CYP1A1 and CYP17A1 were greatly higher in the exposure group than the control group (

0.05)

while treatment of AhR inhibitor CH223191 resulted in notably down-regulated the expression of the 2 proteins (

0.05).

Conclusion

BkF induces the activation of AhR receptors

then causes elevated expression of downstream genes CYP1A1 and CYP17A1 as well as increased intracellular ROS

and finally leads to oxidative stress and genetic damage in GC-2 cells.

关键词

Keywords

references

PATEL A B, SHAIKH S, JAIN K R, et al . Polycyclic aromatic hydrocarbons: sources, toxicity, and remediation approaches[J ] . Front Microbiol, 2020, 11: 562813. DOI: 10.3389/fmicb.2020.562813.

WELLS P G, MCCALLUM G P, LAM K C H, et al . Oxidative DNA damage and repair in teratogenesis and neurodevelopmental deficits[J ] . Birth Defects Res C Embryo Today, 2010, 90(2): 103-109. DOI: 10.1002/bdrc.20177.

RAI P K. Impacts of particulate matter pollution on plants: implications for environmental biomonitoring[J]. Ecotoxicol Environ Saf, 2016, 129: 120-136. DOI: 10.1016/j.ecoenv.2016.03.012.

WANG L L, HU W Y, XIA Y K, et al . Associations between urinary polycyclic aromatic hydrocarbon metabolites and serum testosterone in US adult males: national Health and nutrition examination survey 2011-2012[J ] . Environ Sci Pollut Res Int, 2017, 24(8): 7607-7616. DOI: 10.1007/s11356-017-8407-9.

YANG P, WANG Y X, SUN L, et al . Urinary metabolites of polycyclic aromatic hydrocarbons, sperm DNA damage and spermatozoa apoptosis[J ] . J Hazard Mater, 2017, 329: 241-248. DOI: 10.1016/j.jhazmat.2017.01.053.

CHEN Q, WANG F R, YANG H, et al . Exposure to fine particulate matter-bound polycyclic aromatic hydrocarbons, male semen quality, and reproductive hormones: the MARCHS study[J ] . Environ Pollut, 2021, 280: 116883. DOI: 10.1016/j.envpol.2021.116883.

王芙蓉, 周妮娅, 陈卿, 等. 重庆市大气细颗粒物中多环芳烃组分与男性精子DNA损伤的关联研究[J]. 第三军医大学学报, 2021, 43(1): 17-24. DOI: 10.16016/j.1000-5404.202009206.

WANG F R, ZHOU N Y, CHEN Q, et al . Association between fine particulate matter bound polycyclic aromatic hydrocarbons and DNA damage of male sperm in Chongqing[J ] . J Third Mil Med Univ, 2021, 43(1): 17-24. DOI: 10.16016/j.1000-5404.202009206.

梁继仁, 朱鸿雁, 李翠珍, 等. 大鼠新生期苯并芘暴露对睾丸抗氧化酶活性和精子生成量的影响[J]. 环境与职业医学, 2012, 29(5): 290-293. DOI: 10.13213/j.cnki.jeom. 2012.05.008.

LIANG J R, ZHU H Y, LI C Z, et al . Effects of benzo(a)Pyrene exposure on testicular antioxidant enzyme activities in neonatal SD rats[J ] . J Environ Occup Med, 2012, 29(5): 290-293. DOI: 10.13213/j.cnki.jeom.2012. 05.008.

杨望, 邹鹏, 张国伟, 等. BPDE损伤线粒体抑制小鼠睾丸间质细胞睾酮生成[J]. 第三军医大学学报, 2019, 41(6): 529-535. DOI: 10.16016/j.1000-5404.201901001.

YANG W, ZOU P, ZHANG G W, et al . Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells[J ] . J Third Mil Med Univ, 2019, 41(6): 529-535. DOI: 10.16016/j. 1000-5404.201901001.

李琼, 陈燊, 张海燕, 等. 利用Tox21通路数据库评估石家庄市大气细颗粒物中多环芳烃激活毒性通路的风险[J]. 癌变·畸变·突变, 2020, 32(2): 112-117. DOI: 10.3969/j.issn.1004-616x.2020.02.006.

LI Q, CHEN S, ZHANG H Y, et al . Risk assessment of fine particulate matter exposure using toxicity pathways of human lung cells[J ] . Carcinog Teratog Mutagen, 2020, 32(2): 112-117. DOI: 10.3969/j.issn.1004-616x. 2020.02.006.

IZAWA H, KOHARA M, WATANABE G, et al . Effects of diesel exhaust particles on the male reproductive system in strains of mice with different aryl hydrocarbon receptor responsiveness[J ] . J Reprod Dev, 2007, 53(6): 1191-1197. DOI: 10.1262/jrd.19114.

魏凤华, 张俊江, 夏普, 等. 类二噁英物质及芳香烃受体(AhR)介导的有害结局路径(AOP)研究进展[J]. 生态毒理学报, 2016, 11(1): 37-51. DOI: 10.7524/AJE.1673-5897.20150412001.

WEI F H, ZHANG J J, XIA P, et al . Research progress on dioxin-like compounds and AhR-mediated adverse outcome pathway (AOP)[J ] . Asian J Ecotoxicol, 2016, 11(1): 37-51. DOI: 10.7524/AJE.1673-5897.20150412001.

TONG B, YUAN X S, DOU Y N, et al . Norisoboldine, an isoquinoline alkaloid, acts as an aryl hydrocarbon receptor ligand to induce intestinal Treg cells and thereby attenuate arthritis[J ] . Int J Biochem Cell Biol, 2016, 75: 63-73. DOI: 10.1016/j.biocel.2016.03.014.

薄军, 吴世军, 李裕红, 等. 苯并[a]芘(BaP)对真鲷细胞色素P450和芳香烃受体基因表达的影响[J]. 中山大学学报(自然科学版), 2010, 49(3): 93-97.

BO J, WU S J, LI Y H, et al . The effects of benzo[a ] pyrene(BaP) exposure on the CYP1A1 mRNA and AhR2 mRNA expression of red seabream (Pagrus major)[J ] . Acta Sci Nat Univ Sunyatseni, 2010, 49(3): 93-97.

ZANGAR R C, DAVYDOV D R, VERMA S. Mechanisms that regulate production of reactive oxygen species by cytochrome P450[J]. Toxicol Appl Pharmacol, 2004, 199(3): 316-331. DOI: 10.1016/j.taap.2004.01.018.

KOPF P G, WALKER M K. 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin increases reactive oxygen species production in human endothelial cells via induction of cytochrome P4501A1[J]. Toxicol Appl Pharmacol, 2010, 245(1): 91-99. DOI: 10.1016/j.taap.2010.02.007.

任斐. AHR介导氧化应激参与PM2.5心脏发育毒性及白藜芦醇的保护作用[D]. 苏州: 苏州大学, 2020.

REN F. AHR-mediated oxidative stress contributes to cardiac developmental toxicity of PM2.5 and the protective effect of resveratrol[D]. Suzhou: SooChow University, 2020.

IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. Some non-heterocyclic polycyclic aromatic hydrocarbons and some related exposures[J]. IARC Monogr Eval Carcinog Risks Hum, 2010, 92: 1-853.

任加云, 潘鲁青, 苗晶晶. 苯并(a)芘和苯并(k)荧蒽混合物对栉孔扇贝毒理学指标的影响[J]. 环境科学学报, 2006, 26(7): 1180-1186. DOI: 10.13671/j.hjkxxb.2006. 07.023.

REN J Y, PAN L Q, MIAO J J. Effects of benzo(a)Pyrene and benzo(k)fluoranthene mixture on the toxicology parameter of scallop Chlamys farreri[J]. Acta Sci Circumstantiae, 2006, 26(7): 1180-1186. DOI: 10.13671/j.hjkxxb.2006.07.023.

NEBERT D W, SHI Z Q, GÁLVEZ-PERALTA M, et al . Oral benzo[a ] Pyrene: understanding pharmacokinetics, detoxication, and consequences: Cyp1 knockout mouse lines as a paradigm[J ] . Mol Pharmacol, 2013, 84(3): 304-313. DOI: 10.1124/mol.113.086637.

HEAD J A, JEFFERY R W, FARMAHIN R, et al . Potency of polycyclic aromatic hydrocarbons (PAHs) for induction of ethoxyresorufin-O-deethylase (EROD) activity in hepatocyte cultures from chicken, Pekin duck,and greater scaup[J ] . Environ Sci Technol, 2015, 49(6): 3787-3794. DOI: 10.1021/acs.est.5b00125.

SPINK D C, WU S J, SPINK B C, et al . Induction of CYP1A1 and CYP1B1 by benzo(k)fluoranthene and benzo(a)Pyrene in T-47D human breast cancer cells: roles of PAH interactions and PAH metabolites[J ] . Toxicol Appl Pharmacol, 2008, 226(3): 213-224. DOI: 10.1016/j.taap. 2007.08.024.

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