[1]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.
 JIANG Fan,ZHANG Guowei,AO Lin,et al.BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells[J].J Third Mil Med Univ,2017,39(23):2323-2328.
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BPDE诱导GC-1细胞凋亡及褪黑素的保护作用(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
39卷
期数:
2017年第23期
页码:
2323-2328
栏目:
公共卫生与预防医学
出版日期:
2017-12-15

文章信息/Info

Title:
BPDE induces apoptosis while melatonin protects in spermatogonial GC-1 cells
作者:
姜帆张国伟敖琳曹佳
第三军医大学军事预防医学院毒理学研究所
Author(s):
JIANG Fan ZHANG Guowei AO Lin CAO Jia

Institute of Toxicology, College of Military Preventive Medicine, Third Military Medical University, Chongqing, 400038, China

关键词:
78-二羟-910-环氧苯并[a]芘褪黑素活性氧细胞凋亡
Keywords:
benzo[a]pyrene-78-diol-910-epoxide melatonin reactive oxygen species apoptosis
分类号:
R321.1; R977.1; R996
文献标志码:
A
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. Conclusion     BPDE induces mitochondria-dependent apoptosis and oxidative stress in GC-1 cells, while melatonin protects the cells in the process.

更新日期/Last Update: 2017-12-12