浏览全部资源
扫码关注微信
BPDE损伤线粒体抑制小鼠睾丸间质细胞睾酮生成
Role of mitochondrial damage in BPDE-induced inhibition of testosterone production in mouse Leydig cells
- 第三军医大学学报 2019年41卷第6期 页码：529-535
- 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：
[曹佳，E-mail：caojia1962@126.com CAO Jia, E-mail:caojia1962@126.com
- 基金信息：国家重点研发计划(2017YFC1600202)
DOI：10.16016/j.1000-5404.201901001
中图分类号： Q579.11;R12;R322.64收稿：2019-01-01，
修回：2019-02-07，
纸质出版：2019-03-30
- 稿件说明：
移动端阅览
杨望, 邹鹏, 张国伟, 等. BPDE损伤线粒体抑制小鼠睾丸间质细胞睾酮生成[J]. 第三军医大学学报, 2019,41(6):529-535.
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.
杨望, 邹鹏, 张国伟, 等. BPDE损伤线粒体抑制小鼠睾丸间质细胞睾酮生成[J]. 第三军医大学学报, 2019,41(6):529-535. DOI： 10.16016/j.1000-5404.201901001.
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
2
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
2
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
2
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%(
P
<
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(
P
<
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(
P
<
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(
P
<
0.05)。
结论
2
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
2
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
2
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
25
50
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
2
BPDE at the doses of 25
50
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 (
P
<
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 (
P
<
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 (
P
<
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 (
P
<
0.05).
Conclusion
2
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
HAN X, ZHOU N Y, CUI Z H, et al . Association between urinary polycyclic aromatic hydrocarbon metabolites and sperm DNA damage:A population study in Chongqing, China[J ] . Environ Health Perspect, 2011, 119(5):652-657. DOI:10.1289/ehp.1002340.
HSU P C, CHEN I Y, PAN C H, et al . Sperm DNA damage correlates with polycyclic aromatic hydrocarbons biomarker in coke-oven workers[J ] . Int Arch Occup Environ Health, 2006, 79(5):349-356. DOI:10.1007/s00420-005-0066-3.
XIA Y K, HAN Y, ZHU P F, et al . Relation between urinary metabolites of polycyclic aromatic hydrocarbons and human semen quality[J ] . Environ Sci Technol, 2009, 43(12):4567-4573.
ARCHIBONG A E, RAMESH A, NIAZ M S, et al . Effects of benzo(a)pyrene on intra-testicular function in F-344 rats[J ] . Int J Environ Res Public Health, 2008, 5(1):32-40.
INYANG F, RAMESH A, KOPS OMBUT P, et al . Disruption of testicular steroidogenesis and epididymal function by inhaled benzo(a)pyrene[J ] . Reprod Toxicol, 2003, 17(5):527-537.
CHUNG J Y, KIM Y J, KIM J Y, et al . Benzo[a ] pyrene reduces testosterone production in rat Leydig cells via a direct disturbance of testicular steroidogenic machinery[J ] . Environ Health Perspect, 2011, 119(11):1569-1574. DOI:10.1289/ehp.1003391..
ANNAMALAI J,NAMASIVAYAM V. Endocrine disrupting chemicals in the atmosphere:Their effects on humans and wildlife[J]. Environ Int, 2015, 76:78-97. DOI:10.1016/j.envint.2014.12.006.
KULLER L H, GARFINKEL L, CORREA P, et al . Contribution of passive smoking to respiratory cancer[J ] . Environ Health Perspect, 1986, 70:57-69. DOI:10.1289/ehp.867057.
GLANCY B, HARTNELL L M, MALIDE D, et al . Mitochondrial reticulum for cellular energy distribution in muscle[J ] . Nature, 2015, 523(7562):617-620. DOI:10.1038/nature14614.
FA S, POGRMIC-MAJKIC K, SAMARDZIJA D, et al . HBCDD-induced sustained reduction in mitochondrial membrane potential, ATP and steroido genesis in peripubertal rat Leydig cells[J ] . Toxicol Appl Pharmacol, 2015, 282(1):20-29. DOI:10.1016/j.taap.2014.11.001.
HALES D B. Testicular macrophage modulation of Leydig cell steroidogenesis[J]. J Reprod Immunol, 2002, 57(1/2):3-18.
WARREN J S, TRACY C M, MILLER M R, et al . Histone methyltransferase Smyd1 regulates mitochondrial energetics in the heart[J ] . Proc Natl Acad Sci USA, 2018, 115(33):E7871-E7880. DOI:10.1073/pnas.1800680115.
ITAMI N, SHIRATSUKI S, SHIRASUNA K, et al . Mitochondrial biogenesis and degradation are induced by CCCP treatment of porcine oocytes[J ] . Reproduction, 2015, 150(2):97-104. DOI:10.1530/REP-15-0037.
BANSAL S, LEU A N, GONZALEZ F J, et al . Mitochondrial targeting of cytochrome P450 (CYP) 1B1 and its role in polycyclic aromatic hydrocarbon-induced mitochondrial dysfunction[J ] . J Biol Chem, 2014, 289(14):9936-9951. DOI:10.1074/jbc.M113.525659.
MARQUES V B, FARIA R A, DOS SANTOS L. Overview of the pathophysiological implications of organotins on the endocrine system[J]. Front Endocrinol (Lausanne), 2018, 9:101. DOI:10.3389/fendo.2018.00101.
WONG J Y Y, HU W, DOWNWARD G S, et al . Personal exposure to fine particulate matter and benzo[a ] pyrene from indoor air pollution and leukocyte mitochondrial DNA copy number in rural China[J ] . Carcinogenesis, 2017, 38(9):893-899. DOI:10.1093/carcin/bgx068.
LING X, ZHANG G W, SUN L, et al . Polycyclic aromatic hydrocarbons exposure decreased sperm mitochondrial DNA copy number:A cross-sectional study (MARHCS) in Chongqing, China[J ] . Environ Pollut, 2017, 220(Pt A):680-687. DOI:10.1016/j.envpol.2016.10.026.
0
浏览量
0
下载量
0
CSCD
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
