Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells

Wang YANG; Peng ZOU; Guowei ZHANG; Xi LING; Jinyi LIU; Lin AO; Jia CAO

doi:10.16016/j.1000-5404.201901001

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Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells

Basic Medicine | 更新时间：2026-01-09

- Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells
- Journal of Third Military Medical University Vol. 41, Issue 6, Pages: 529-535(2019)
- 作者机构：
  
  1.400038 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
  2.400038 重庆，陆军军医大学(第三军医大学)军事预防医学系:环境卫生学教研室
- 作者简介：
- 基金信息：
  
  the National Key R&D Program of China(2017YFC1600202)
- DOI：10.16016/j.1000-5404.201901001
  CLC： Q579.11;R12;R322.64
- Received：01 January 2019，
  
  Revised：2019-02-07，
  
  Published：30 March 2019
- 稿件说明：
移动端阅览
Wang YANG, Peng ZOU, Guowei ZHANG, et al. Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells[J]. Journal of Third Military Medical University, 2019, 41(6): 529-535.
DOI：

Wang YANG, Peng ZOU, Guowei ZHANG, et al. Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells[J]. Journal of Third Military Medical University, 2019, 41(6): 529-535. DOI： 10.16016/j.1000-5404.201901001.

摘要

目的

观察苯并[a]芘-7

8-二醇-9

10-环氧化物(benzo[a]pyrene-r-7

t-8-dihydrodiol-t-9

10-epoxide(±)

(anti)

BPDE)对睾丸间质细胞睾酮生成的影响，并围绕线粒体损伤探讨其作用机制。

方法

使用不同浓度BPDE对小鼠睾丸间质细胞株TM3进行染毒，48 h后采用台盼蓝染色法检测TM3细胞的细胞活率。采用不引起细胞明显死亡的剂量进行后续实验。分别使用0(对照组)、25、50、100 nmol/L的BPDE处理细胞，48 h后使用睾酮ELISA试剂盒检测培养基中睾酮浓度，透射电镜观察细胞线粒体超微结构，JC-1探针检测细胞线粒体膜电位，ATP检测试剂盒检测细胞内ATP水平，荧光定量PCR检测线粒体DNA拷贝数，Western blot检测PGC-1α、COX Ⅳ、StAR和TSPO的表达。采用线粒体保护剂ZLN005对细胞进行预处理，检测100 nmol/L BPDE染毒对上述指标的影响。

结果

与对照组比较，25、50、100 nmol/L BPDE不造成TM3细胞活率的明显改变；50和100 nmol/L BPDE处理可分别导致细胞睾酮水平降低21.15%和34.96%(

0.01)，并造成线粒体结构功能的改变，包括线粒体超微结构的改变，线粒体膜电位、细胞ATP水平和线粒体DNA拷贝数降低(

0.05)。同时线粒体生物合成相关蛋白PGC-1α、线粒体功能相关蛋白COX Ⅳ、睾酮合成关键蛋白StAR和TSPO的表达均出现剂量依赖性降低。与100 nmol/L BPDE单独染毒组比较，线粒体保护剂ZLN005预处理组细胞睾酮水平明显升高(

0.05)，同时线粒体相关指标，包括线粒体膜电位、细胞ATP水平、线粒体DNA拷贝数、线粒体生物生成和睾酮合成关键蛋白的表达均显著上升(

0.05)。

结论

BPDE可造成TM3细胞线粒体损伤及睾酮合成水平降低，增加线粒体生物合成和保护线粒体功能可缓解BPDE所致的睾酮水平降低。

Abstract

Objective

To observe the effect of benzo[a]pyrene-r-7

t-8-dihydrodiol-t-9

10-epoxide(±)

(anti) (BPDE) on testosterone synthesis from mouse Leydig cells

and investigate the potential mechanism involved mitochondrial damage.

Methods

TM3 mouse Leydig cells were treated with various doses of BPDE for 48 h. Cell viability was assessed by trypan blue assay. Then the doses under the lethal dose were selected according to results of cell viability. After TM3 cells were treated with 0

or 100 nmol/L BPDE for 48 h

testosterone concentration in the medium

mitochondrial ultrastructure of TM3 cells

mitochondrial membrane potential

ATP level

mitochondrial DNA copy number

and expression levels of PGC-1α

COX Ⅳ

StAR and TSPO were measured through ELISA kit for testosterone

transmission electron microscopy

JC-1 probe

ATP detection kit

quantitative PCR and Western blotting

respectively. Furthermore

TM3 cells were pre-treated with 5 μmol/L ZLN005

a regulator to protect mitochondrial function and stimulate mitochondrial biogenesis. Then

after 24 h

these pre-treated cells were treated with 100 nmol/L BPDE for 48 h

the indicators above were determined in the same ways.

Results

BPDE at the doses of 25

100 nmol/L had no significantly effect on cell viability of TM3 cells. Compared to control cells

50 and 100 nmol/L BPDE reduced testosterone synthesis by 21.15% and 34.96%

respectively (

0.01)

and also induced mitochondrial structure and function

including alterations of mitochondrial ultrastructure

and appreciable decreases in mitochondrial membrane potential

cellular ATP production and mitochondrial DNA copy number (

0.05). What's more

the levels of proteins related to mitochondrial function

COX Ⅳ and to mitochondrial biogenesis

PGC-1α

and the key proteins for testosterone synthesis

StAR and TSPO

were all reduced in a dose-dependent manner. Compared to TM3 cells treated with 100 nmol/L BPDE alone

pre-treatment with 5 μmol/L ZLN005 for 24 h significantly elevated testosterone level in the medium (

0.05). Notable restores of the indicators above were observed in the TM3 cells pre-treated with ZLN005

including mitochondrial membrane potential

cellular ATP production

mitochondrial DNA copy number

and protein levels of COX Ⅳ

PGC-1α

StAR and TSPO (

0.05).

Conclusion

BPDE induces reduction of testosterone synthesis

via

mitochondrial compromise. Protecting mitochondria could alleviate BPDE-induced mitochondrial damage and reduction of testosterone synthesis.

关键词

Keywords

references

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