BPDE诱导GC-1细胞凋亡及褪黑素的保护作用

姜帆; 张国伟; 敖琳; 曹佳

doi:10.16016/j.1000-5404.201708180

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BPDE诱导GC-1细胞凋亡及褪黑素的保护作用

公共卫生与预防医学 | 更新时间：2026-01-09

- BPDE诱导GC-1细胞凋亡及褪黑素的保护作用
- BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells
- 第三军医大学学报 2017年39卷第23期页码：2323-2328
- 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：
  
  [曹佳，E-mail：caojia1962@126.com CAO Jia, E-mail: caojia1962@126.com
- 基金信息：
  
  国家自然科学基金重点项目(81130051)
- DOI：10.16016/j.1000-5404.201708180
  中图分类号： R321.1;R977.1;R996
- 收稿：2017-08-23，
  
  修回：2017-09-20，
  
  纸质出版：2017-12-15
- 稿件说明：
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姜帆, 张国伟, 敖琳, 等. BPDE诱导GC-1细胞凋亡及褪黑素的保护作用[J]. 第三军医大学学报, 2017,39(23):2323-2328.

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.
姜帆, 张国伟, 敖琳, 等. BPDE诱导GC-1细胞凋亡及褪黑素的保护作用[J]. 第三军医大学学报, 2017,39(23):2323-2328. DOI： 10.16016/j.1000-5404.201708180.

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

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