PARP-1在鱼藤酮诱导PC12细胞线粒体DNA损伤中的作用

肖靖淞; 侯贵钟; 赛燕; 张爱华; 曹佳

doi:10.16016/j.1000-5404.201812113

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PARP-1在鱼藤酮诱导PC12细胞线粒体DNA损伤中的作用

基础医学 | 更新时间：2026-03-02

- PARP-1在鱼藤酮诱导PC12细胞线粒体DNA损伤中的作用
- Role of PARP-1 in mitochondroal DNA damage induced by rotenone in PC12 cells
- 第三军医大学学报 2019年41卷第8期页码：778-785
- 作者机构：
  
  1..550025 贵阳，贵州医科大学公共卫生学院卫生毒理学教研室Department of Toxicology, College of Public Health, Guizhou Medical University, Guiyang, Guizhou Province, 550025
  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Institute of Toxicology, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, China
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目(81473006)
- DOI：10.16016/j.1000-5404.201812113
  中图分类号： R363.21;R732.502;R977.3
- 收稿：2018-12-13，
  
  纸质出版：2019-04-30
- 稿件说明：
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肖靖淞, 侯贵钟, 赛燕, 等. PARP-1在鱼藤酮诱导PC12细胞线粒体DNA损伤中的作用[J]. 第三军医大学学报, 2019,41(8):778-785.

Jingsong XIAO, Guizhong HOU, Yan SAI, et al. Role of PARP-1 in mitochondroal DNA damage induced by rotenone in PC12 cells[J]. Journal of Third Military Medical University, 2019, 41(8): 778-785.
肖靖淞, 侯贵钟, 赛燕, 等. PARP-1在鱼藤酮诱导PC12细胞线粒体DNA损伤中的作用[J]. 第三军医大学学报, 2019,41(8):778-785. DOI： 10.16016/j.1000-5404.201812113.

Jingsong XIAO, Guizhong HOU, Yan SAI, et al. Role of PARP-1 in mitochondroal DNA damage induced by rotenone in PC12 cells[J]. Journal of Third Military Medical University, 2019, 41(8): 778-785. DOI： 10.16016/j.1000-5404.201812113.

摘要

目的

探讨PARP-1在鱼藤酮(rotenone)诱导大鼠PC12细胞线粒体DNA(mtDNA)损伤中的作用。

方法

不同浓度(0、0.1、0.5、1.0、1.5 μmol/L)鱼藤酮处理PC12细胞，TaqMan探针法PCR检测mtDNA缺失，qPCR法检测mtDNA拷贝数，Western blot检测PARP-1蛋白在PC12细胞内和线粒体内的表达水平。PARP-1选择性抑制剂PJ34预处理鱼藤酮染毒的PC12细胞，采用CCK-8法检测细胞增殖活力，TaqMan探针法PCR检测mtDNA缺失，qPCR法检测mtDNA拷贝数，四甲基罗丹明乙酯(TMRM)染色检测线粒体膜电位，DCFH-DA探针检测细胞内活性氧(reactive oxygen species，ROS)水平。

结果

TaqMan探针法PCR和qPCR法检测结果显示，与对照组(0 μmol/L鱼藤酮)比较，鱼藤酮处理可引起PC12细胞线粒体DNA缺失增加和线粒体DNA拷贝数降低(

0.05)。同时，细胞和线粒体内的PARP-1蛋白表达被激活，Western bolt检测结果显示，PARP-1蛋白表达水平随鱼藤酮浓度增加而增加。与鱼藤酮单独处理组比较，PJ34预处理可提高鱼藤酮染毒引起的PC12细胞的活力下降(

0.05)，并降低线粒体DNA缺失和提高线粒体DNA拷贝数水平(

0.05)。TMRM染色结果显示，PJ34预处理后线粒体膜电位水平提高(

0.05)；DCFH-DA探针检测结果显示，PJ34预处理后细胞内ROS水平下降(

0.05)。

结论

鱼藤酮可诱导PC12细胞线粒体DNA的损伤，激活PC12细胞和线粒体内PARP-1蛋白的表达，PARP-1选择性抑制剂PJ34对鱼藤酮诱导的多巴胺神经元PC12细胞线粒体DNA的损伤具有保护作用。

Abstract

Objective

To explore the effect of poly (ADP-ribose) polymerase-1 (PARP-1) on mitochondrial DNA (mtDNA) damage induced by rotenone in rat adrenal medulla pheochromocytoma PC12 cells.

Methods

PC12 cells were treated with various dose of rotenone. The mtDNA common deletion (CDs) was determined by TaqMan assay

and mtDNA copy number was detected by quantitative polymerase chain reaction (qPCR). Western blotting was performed to detect the protein levels of PARP-1 in whole cell and mitochondria. After the rotenone exposed PC12 cells were treated with PJ34 (PARP-1 selective inhibitor)

cell viability was detected by CCK-8 assay

mitochondrial membrane potential was examined by TMRM staining

intracellular reactive oxygen species (ROS) level was tested by 2

7-dichlorofluorescin diacetate (DCFH-DA) probe.

Results

TaqMan and qPCR assays showed that rotenone treatment induced increased mtDNA deletion and decreased mtDNA copy number in PC12 cells when compared to control cells (

0.05). Meanwhile

PARP-1 was activated by rotenone treatment in the whole cells and mitochondria

and Western boltting showed that the protein level of PARP-1 was upregulated in a dose-dependent manner. Furthermore

compared with the rotenone group

PJ34 pretreatment enhanced the cell viability significantly in PC12 cells (

0.05)

abolished mtDNA CDs and increased mtDNA copy number (

0.05). Additionally

PJ34 pretreatment caused the improvement of mitochondrial membrane potential and reduced ROS production when compared with the rotenone cells(

0.05).

Conclusion

Rotenone induces mitochondrial DNA damage

which further activates the expression of PARP-1 protein

especially mitochondria in PC12 cells. And PJ34

as the PARP-1 selective inhibitor has a protective effect on rotenone-induced mitochondrial DNA damage in PC12 cells.

关键词

Keywords

references

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