BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells

Fan JIANG; Guowei ZHANG; Lin AO; Jia CAO

doi:10.16016/j.1000-5404.201708180

您当前的位置：

首页 >

文章列表页 >

BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells

Public Health and Preventive Medicine | 更新时间：2026-01-09

- BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells
- Journal of Third Military Medical University Vol. 39, Issue 23, Pages: 2323-2328(2017)
- 作者机构：
  
  400038 重庆, 第三军医大学军事预防医学院毒理学研究所
- 作者简介：
- 基金信息：
  
  the Key Program of National Natural Science Foundation of China(81130051)
- DOI：10.16016/j.1000-5404.201708180
  CLC： R321.1;R977.1;R996
- Received：23 August 2017，
  
  Revised：2017-09-20，
  
  Published：15 December 2017
- 稿件说明：
移动端阅览
Fan JIANG, Guowei ZHANG, Lin AO, et al. BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells[J]. Journal of Third Military Medical University, 2017, 39(23): 2323-2328.
DOI：

Fan JIANG, Guowei ZHANG, Lin AO, et al. BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells[J]. Journal of Third Military Medical University, 2017, 39(23): 2323-2328. DOI： 10.16016/j.1000-5404.201708180.

摘要

目的

探讨7

8-二羟-9

10-环氧苯并[a]芘(benzo[a]pyrene-7

8-diol-9

10-epoxide

BPDE)对小鼠精原细胞株GC-1的细胞毒性机制及褪黑素的保护作用。

方法

不同浓度BPDE处理GC-1细胞，采用Annexin V/PI染色及流式细胞仪检测细胞凋亡，JC-1探针染色检测线粒体膜电位，Western blot检测细胞质Cyt C及细胞总蛋白中caspase-9/3的活化水平，DCFH-DA探针及流式细胞仪检测细胞内的活性氧(reactive oxygen species

ROS)水平。褪黑素预处理细胞后再进行BPDE染毒，采用上述方法检测褪黑素对BPDE细胞毒性的影响。

结果

BPDE可提高GC-1细胞凋亡率，存在剂量-效应关系。同时，BPDE可降低细胞线粒体膜电位，促进线粒体Cyt C的释放，提高细胞中caspase-9和caspase-3蛋白的活化水平，并诱导细胞中ROS水平升高。褪黑素预处理则可降低BPDE诱导的细胞凋亡，抑制Cyt C的释放及caspase-9/3的活化，并激活Nrf-2/ARE抗氧化通路，降低细胞ROS水平。

结论

BPDE可诱导小鼠精原细胞GC-1线粒体途径细胞凋亡和氧化应激，而褪黑素在这一过程中起保护作用。

Abstract

Objective

To explore the mechanism of benzo[a]pyrene-7

8-diol-9

10-epoxide (BPDE)-induced cytotoxicity in mouse spermatogonial GC-1 cells and the protective role of melatonin in the process.

Methods

GC-1 cells were treated with various doses of BPDE. Then

cell apoptosis was detected using Annexin V/PI staining and flow cytometry analysis

and mitochondrial membrane potential was examined by JC-1 staining. After the proteins in the cytoplasm and whole cells were extracted separately

the expression of Cyt C

active caspase-9 and active caspase-3 was measured using Western blot assay. 2

7-dichloro-fluorescin diacetate (DCFH-DA) probe and flow cytometry were used to measure intracellular reactive oxygen species (ROS) level. The effect of melatonin pretreatment on the induced cytotoxicity in GC-1 cells was also studied.

Results

BPDE induced cell apoptosis in GC-1 cells in a dose-dependent manner. BPDE treatment also resulted in decrease in the mitochondrial membrane potential

enhancement in the release of cytoplasmic Cyt C from mitochondria

increases in the expression levels of active caspase-9 and -3

and elevation in intracellular ROS level in the GC-1 cells. Furthermore

pretreatment of melatonin significantly inhibited BPDE-induced apoptosis

release of cytoplasmic Cyt C and activation of caspase-9

and activate the Nrf-2/ARE antioxidant pathway and abolished the increase of ROS in GC-1 cells.

Conclusions

BPDE induces mitochondria-dependent apoptosis and oxidative stress in GC-1 cells

while melatonin protects the cells in the process.

关键词

Keywords

references

CONTI G O, CALOGERO A E, GIACONE F, et al . B(a)P adduct levels and fertility: A cross-sectional study in a Sicilian population[J ] . Mol Med Rep, 2017, 15(5): 3398-3404. DOI:10.3892/mmr.2017.6396.

ARAFA H M, ALY H A, ABD-ELLAH M F, et al . Hesperidin attenuates benzo[alpha ] pyrene-induced testicular toxicity in rats via regulation of oxidant/antioxidant balance[J ] . Toxicol Ind Health, 2009, 25(6): 417-427. DOI:10.1177/0748233709106624..

O'BRIEN J M, BEAL M A, YAUK C L, et al . Benzo(a)pyrene is mutagenic in mouse spermatogonial stem cells and dividing spermatogonia[J ] . Toxicol Sci, 2016, 152(2): 363-371. DOI:10.1093/toxsci/kfw088.

RANJIT S, MIDDE N M, SINHA N, et al . Effect of polyaryl hydrocarbons on cytotoxicity in monocytic cells: potential role of cytochromes P450 and oxidative stress pathways[J ] . PLoS ONE, 2016, 11(9): e0163827. DOI:10.1371/journal.pone.0163827.

XIAO H, RAWAL M, HAHM E R, et al . Benzo[a ] pyrene-7, 8-diol-9, 10-epoxide causes caspase-mediated apoptosis in H460 human lung cancer cell line[J ] . Cell Cycle, 2007, 6(22): 2826-2834. DOI:10.4161/cc.6.22.4891.

张天宝, 侯鹏高.褪黑素生理活性及其作用机制的研究进展[J].齐齐哈尔医学院学报, 2015, 36 (11):1671-1673.

ZHANG T B, HOU G P. Progress on physiological activities of melatonin and its mechanisms [J]. J Qiqihar Med Coll, 2015, 36 (11): 1671-1673.

SUZEN S. Melatonin and synthetic analogs as antioxidants[J]. Curr Drug Deliv, 2013, 10(1): 71-75. DOI:10.2174/1567201811310010013.

WASEEM M, SAHU U, SALMAN M, et al . Melatonin pre-treatment mitigates SHSY-5Y cells against oxaliplatin induced mitochondrial stress and apoptotic cell death[J ] . PLoS ONE, 2017, 1 2(7): e0180953. DOI:10.1371/journal.pone.0180953.

CARLSEN E, GIWERCMAN A, KEIDING N, et al . Evidence for decreasing quality of semen during past 50 years[J ] . BMJ, 1992, 305(6854): 609-613. DOI:10.1136/bmj.305.6854.609.

张树成, 王弘毅, 王介东. 1981~1996年我国有生育力男性精液质量的变化分析[J].生殖与避孕, 1999, 19(1): 27-33.

ZHANG S C, WANG H Y, WANG J D. Analysis of change in sperm quality of Chinese fertile men during 1981-1996 [J]. Reprod Contracep, 1999, 19 (1):27-33.

BARAZANI Y, KATZ B F, NAGLER H M, et al . Lifestyle, environment, and male reproductive health[J ] . Urol Clin North Am, 2014, 41(1): 55-66. DOI:10.1016/j.ucl.2013.08.017.

SANG H, ZHANG L, LI J. Anti-benzopyrene-7, 8-diol-9, 10-epoxide induces apoptosis via mitochondrial pathway in human bronchiolar epithelium cells independent of the mitochondria permeability transition pore[J ] . Food Chem Toxicol, 2012, 50(7): 2417-2423. DOI:10.1016/j.fct.2012.04.041.

LI Y, LI S, ZHOU Y, et al . Melatonin for the prevention and treatment of cancer[J ] . Oncotarget, 2017, 8(24): 3 9896-39921. DOI:10.18632/oncotarget.16379.

XIE Z, CHEN F, LI W A, et al . A review of sleep disorders and melatonin[J ] . Neurol Res, 2017, 39(6): 559-565. DOI:10.1080/01616412.2017.1315864.

MURAWSKA-CIAŁOWICZ E, JETHON Z, MAGDALAN J, et al . Effects of melatonin on lipid peroxidation and antioxidative enzyme activities in the liver, kidneys and brain of rats administered with benzo(a)pyrene[J ] . Exp Toxicol Pathol, 2011, 63(1/2): 97-103. DOI:10.1016/j.etp.2009.10.002.

MA Q. Role of nrf2 in oxidative stress and toxicity[J]. Annu Rev Pharmacol Toxicol, 2013, 53: 401-426. DOI:10.1146/annurev-pharmtox-011112-140320.

CHEN L Y, RENN T Y, LIAO W C, et al . Melatonin successfully rescues hippocampal bioenergetics and improves cognitive function following drug intoxication by promoting Nrf2-ARE signaling activity[J ] . J Pineal Res, 2017, 63(2): e12417. DOI:10.1111/jpi.12417.

JANJETOVIC Z, JARRETT S, LEE E, et al . Melatonin and its metabolites protect human melanocytes against UVB-induced damage: Involvement of NRF2-mediated pathways[J ] . Sci Rep, 2017, 7(1): 1274. DOI:10.1038/s41598-017-01305-2.

CAO Z, FANG Y, LU Y, et al . Melatonin alleviates cadmium-induced liver injury by inhibiting the TXNIP-NLRP3 inflammasome[J ] . J Pineal Res, 2017, 62(3): e12389. DOI:10.1111/jpi.12389.

JI Y L, WANG H, MENG C, et al . Melatonin alleviates cadmium-induced cellular stress and germ cell apoptosis in testes[J ] . J Pineal Res, 2012, 52(1): 71-79. DOI:10.1111/j.1600-079X.2011.00921.x.

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Palmitic acid increases apoptosis by mitochondrial pathway in hepatocytes with growth hormone deficiency

Protective effects of tea polyphenols on apoptosis and oxidative stress induced by ⁶⁰Coγ radiation in L5178Y cells

Allopurinol inhibits cardiomyocyte apoptosis in rats with myocardial infarction

Effect of cinnamaldehyde on Bax/Bak and apoptosis of vascular endothelial cells in diabetic ulcers

RANKL通过ROS介导HIF-1α下调促进自噬和破骨细胞分化

Related Author

王蕴婷

郑晓雅

陈月

钱文杰

伍询

任伟

李奇慧

唐木涛

Related Institution

Hunan Provincial Key Laboratory of Vascular Biology and Translational Medicine， Hunan University of Chinese Medicine

Department of Burn Wounds and Plastic Surgery， First Affiliated Hospital of Hunan University of Chinese Medicine

Department of Nuclear Medicine， First Affiliated Hospital， Army Medical University （Third Military Medical University）

College of Bioengineering， Chongqing University

⁰