DEHP通过抑制<italic style="font-style: italic">Fto</italic>表达诱导睾丸间质细胞铁死亡

孙凤琼; 张国伟; 王灵巧; 徐桂勇; 郭成威; 孙燕; 杨芮; 张露; 杨光红; 周紫垣; 游明丹

doi:10.16016/j.2097-0927.202404119

您当前的位置：

首页 >

文章列表页 >

DEHP通过抑制Fto表达诱导睾丸间质细胞铁死亡

基础医学 | 更新时间：2026-01-09

- DEHP通过抑制Fto表达诱导睾丸间质细胞铁死亡
- DEHP induces ferroptosis in testicular interstitial cells by inhibiting Fto expression
- 陆军军医大学学报 2024年46卷第21期页码：2369-2382
- 作者机构：
  
  1.561113 贵阳，贵州医科大学公共卫生与健康学院，环境污染与疾病监控教育部重点实验室
  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：
  
  [周紫垣，E-mail: ziyuanzhou@tmmu.edu.cn ZHOU Ziyuan, E-mail: ziyuanzhou@tmmu.edu.cn
  [游明丹，E-mail: mdyou_gzmu@outlook.com YOU Mingdan, E-mail: mdyou_gzmu@outlook.com
- 基金信息：
  
  国家重点研发计划项目(2022YFC2702902)
- DOI：10.16016/j.2097-0927.202404119
  中图分类号： R322.64;R992;R994.6
- 收稿：2024-04-25，
  
  修回：2024-05-19，
  
  纸质出版：2024-11-15
- 稿件说明：
移动端阅览
孙凤琼, 张国伟, 王灵巧, 等. DEHP通过抑制Fto表达诱导睾丸间质细胞铁死亡[J]. 陆军军医大学学报, 2024,46(21):2369-2382.

Fengqiong SUN, Guowei ZHANG, Lingqiao WANG, et al. DEHP induces ferroptosis in testicular interstitial cells by inhibiting Fto expression[J]. Journal of Army Medical University, 2024, 46(21): 2369-2382.
孙凤琼, 张国伟, 王灵巧, 等. DEHP通过抑制Fto表达诱导睾丸间质细胞铁死亡[J]. 陆军军医大学学报, 2024,46(21):2369-2382. DOI： 10.16016/j.2097-0927.202404119.

Fengqiong SUN, Guowei ZHANG, Lingqiao WANG, et al. DEHP induces ferroptosis in testicular interstitial cells by inhibiting Fto expression[J]. Journal of Army Medical University, 2024, 46(21): 2369-2382. DOI： 10.16016/j.2097-0927.202404119.

摘要

目的

探讨邻苯二甲酸二(2-乙基己基)酯(di-2-ethylhexyl phthalate，DEHP)诱导睾丸间质细胞铁死亡中RNA去甲基化酶脂肪量和肥胖相关蛋白(fat mass and obesity-associated protein

FTO)的作用及机制。

方法

40只3周龄C57BL/6雄性小鼠通过随机数字表法分为对照组(玉米油)和3组DEHP染毒组(5、250、500 mg/kg体质量)，连续灌胃35 d；TM3小鼠睾丸间质细胞以0、100、200、400 μmol/L邻苯二甲酸单(2-乙基己基)酯(mono-2-ethylhexy

l phthalate

MEHP)处理24 h，质粒转染构建

Fto

过表达TM3细胞。ELISA检测血清睾酮水平，免疫组化检测睾丸组织中蛋白表达，比色法检测睾丸中Fe

、丙二醛和脂质过氧化物水平。甲基化RNA免疫共沉淀、RT-PCR和Western blot检测N6 -甲基腺嘌呤(N6-methyladenosine

m6A)修饰水平。

结果

250、500 mg/kg DEHP染毒组小鼠的血清睾酮水平显著降低(

＜0.01)，睾丸组织Fe

、丙二醛、脂质过氧化物水平显著升高(

＜0.01)，RNA去甲基化酶FTO、铁死亡相关分子铁蛋白重链1(ferritin heavy chain 1

FTH1)和谷胱甘肽过氧化酶4(glutathione peroxidase 4

GPX4)蛋白水平显著下调(

＜0.05)，转铁蛋白受体(transferrin receptor

TFRC)、膜铁转运蛋白(ferroportin

FPN)、环氧合酶-2(cyclooxygenase-2

COX-2)和酰基辅酶A合成酶长链家族成员4(acyl-CoA synthetase long-chain family member 4

ACSL4)蛋白水平显著上调(

＜0.05)。MEHP处理TM3细胞24 h后，细胞活力下降、胞内活性氧(ROS)含量升高，线粒体膜电位(mitochondrial membrane potential

MMP)显著降低(

＜0.01)，

Fto

的mRNA和蛋白水平均显著下调(

＜0.01)，其余铁死亡相关蛋白的变化也与睾丸组织中趋势一致，提示睾丸间质细胞发生了铁死亡。以铁死亡抑制剂Fer-1干预或过表达

Fto

均能显著抑制MEHP诱导的TM3细胞毒性和铁死亡(

＜0.05)，同时过表达

Fto

使

Gpx

4和

Fth

1 mRNA的m6A修饰水平降低(

＜0.05)。

结论

FTO表达抑制引起

Gpx

4、

Fth

1的m6A修饰异常可能是DEHP诱导睾丸间质细胞铁死亡的机制。

Abstract

Objective

To explore the role and mechanism of RNA demethylase fat mass and obesity-associated protein (FTO) in the ferroptosis in testicular interstitial cells induced by di (2-ethylhexyl) phthalate (DEHP).

Methods

Forty 3-week-old C57BL/6 male mice were randomly divided into a control group (corn oil) and 3 dosed DEHP treatment groups (5

250 and 500 mg/kg)

and received an intragastric infusion of corresponding agents for 35 d

respectively. After mouse testicular interstitial TM3 cells was treated with 0

100

200 and 400 μmol/L mono-2-ethylhexyl phthalate (MEHP) for 24 h

corresponding plasmids were transfected to construct

Fto

overexpressing TM3 cells. S

erum testosterone level was detected by ELISA

expression of testicular proteins was detected with immunohistochemical assay

and contents of Fe

malondialdehyde (MDA) and lipid peroxides in the testicle were detected by colorimetry. Methylated RNA immunoprecipitation

RT-PCR

and Western blotting were used to detect the level of N6-methyladenosine (m6A) modification.

Results

In the mice exposed to 250 and 500 mg/kg DEHP

the serum testosterone level was significantly reduced (

0.01)

contents of Fe

MAD and lipid peroxides in testicular tissue were obviously increased (

0.01)

and protein levels of RNA demethylase FTO

and ferroptosis related molecules ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4) were significantly down-regulated (

0.05)

while those of transferrin receptor (TFRC)

ferroportin (FPN)

cyclooxygenase-2 (COX-2)

and acyl-CoA synthetase long-chain family member 4 (ACSL4) were notably up-regulated (

0.05). MEHP treatment for 24 h resulted in remarkably decreased cell viability in the TM3 cells

increased production of intracellular reactive oxygen species (ROS)

reduced mitochondrial membrane potential (MMP) (

0.01)

down-regulated mRNA and protein levels of

Fto

(

0.01)

and the changes in other ferroptosis related proteins were consistent with the trend in testicular tissue

indicating ferroptosis in testicular interstitial cells. Intervention with ferroptosis inhibitor Fer-1 or overexpression of

Fto

significantly inhibited MEHP-induced toxicity and ferroptosis in TM3 cells (

0.05)

and overexpression of

Fto

reduced the m6A modification of

Gpx

4 and

Fth

1 mRNA (

0.05).

Conclusion

Abnormal m6A modification of

Gpx

4 and

Fth

1 caused by inhibiting FTO expression may be the mechanism of ferroptosis in testicular interstitial cells induced by DEHP.

关键词

Keywords

references

EISE NBERG M L, ESTEVES S C, LAMB D J, et al . Male infertility[J ] . Nature reviews Disease primers, 2023, 9(1): 49. DOI 10.1038/s41572-023-00459-w.

WU H T, ESTILL M S, SHERSHEBNEV A, et al . Preconception urinary phthalate concentrations and sperm DNA methylation profiles among men undergoing IVF treatment: a cross-sectional study[J ] . Hum Reprod, 2017, 32(11): 2159-2169. DOI: 10.1093/humrep/dex283.

ZHAO Y, CHEN M S, WANG J X, et al . Connexin-43 is a promising target for lycopene preventing phthalate-induced spermatogenic disorders[J ] . J Adv Res, 2023, 49: 115-126. DOI: 10.1016/j.jare.2022.09.001.

JIANG X J, STOCKWELL B R, CONRAD M. Ferroptosis: mechanisms, biology and role in disease[J]. Nat Rev Mol Cell Biol, 2021, 22(4): 266-282. DOI: 10.1038/s41580-020-00324-8.

HOSSEINZADEH A, MEHRZADI S, SIAHPOOSH A, et al . The ameliorative effect of ellagic acid on di-(2-ethylhexyl) phthalate-induced testicular structural alterations, oxidative stress, inflammation and sperm damages in adult mice[J ] . Reprod Biol Endocrinol, 2021, 19(1): 146. DOI: 10.1186/s12958-021-00830-0.

ZHAO Y, LI M Z, SHEN Y, et al . Lycopene prevents DEHP-induced leydig cell damage with the Nrf2 antioxidant signaling path way inmice[J ] . J Agric Food Chem, 2020, 68(7): 2031-2040. DOI: 10.1021/acs.jafc.9b06882.

MENG P, ZHANG S S, JIANG X J, et al . Arsenite induces testicular oxidative stress in vivo and in vitro leading to ferroptosis[J ] . Ecotoxicol Environ Saf, 2020, 194: 110360. DOI: 10.1016/j.ecoenv.2020.110360..

WANG J K, ZHANG Z H, SHI F Q, et al . PM 2.5 caused ferroptosis in spermatocyte via overloading iron and disrupting redox homeostasis[J ] . Sci Total Environ, 2023, 872: 162089. DOI: 10.1016/j.scitotenv.2023.162089..

OLUWAYIOSE O A, MARCHO C, WU H T, et al . Paternal preconception phthalate exposure alters sperm methylome and embryonic programming[J ] . Environ Int, 2021, 155: 106693. DOI: 10.1016/j.envint.2021.106693.

ZHANG T, ZHOU Y, LI L, et al . Melatonin protects prepuberal testis from deleterious effects of bisphenol A or diethylhexyl phthalate by preserving H3K9 methylation[J ] . J Pineal Res, 2018, 65(2): e12497. DOI: 10.1111/jpi.12497.

ZHANG J Y, QIU T M, YAO X F, et al . Insights into the role of N6-methyladenosine in ferroptosis[J ] . Biomed Pharmacother, 2023, 165: 115192. DOI: 10.1016/j.biopha.2023.115192.

WANG Z Y, LI H Y, CAI H S, et al . FTO sensitizes oral squamous cell carcinoma to ferroptosis via suppressing ACSL3 and GPX4[J ] . Int J Mol Sci, 2023, 24(22): 16339. DOI: 10.3390/ijms242216339..

JIANG T Y, XIAO Y, ZHOU J F, et al . Arbutin alleviates fatty liver by inhibiting ferroptosis via FTO/SLC7A11 pathway[J ] . Redox Biol, 2023, 68: 102963. DOI: 10.1016/j.redox.2023.102963..

LIN Z, HSU P J, XING X D, et al . Mettl3-/Mettl14-mediated mRNA N 6 -methyladenosine modulates murine spermatogenesis[J ] . Cell Res, 2017, 27(10): 1216-1230. DOI: 10.1038/cr.2017.117..

ZHAO T X, WANG J K, WU Y H, et al . Increased m6A modification of RNA methylation related to the inhibition of demethylase FTO contributes to MEHP-induced Leydig cell injury[J ] . Environ Pollut, 2021, 268(Pt A): 115627. DOI: 10.1016/j.envpol.2020.115627.

YU Z, WANG F, HAN J Y, et al . Opposite effects of high- and low-dose di-(2-ethylhexyl) phthalate (DEHP) exposure on puberty onset, oestrous cycle regularity and hypothalamic kisspeptin expression in female rats[J ] . Reprod Fertil Dev, 2020, 32(6): 610-618. DOI: 10.1071/RD19024.

ZHAO T X, WANG J K, SHEN L J, et al . Increased m6A RNA modification is related to the inhibition of the Nrf2-mediated antioxidant response in di-(2-ethylhexyl) phthalate-induced prepubertal testicular injury[J ] . Environ Pollut, 2020, 259: 113911. DOI: 10.1016/j.envpol.2020.113911.

HA M, GUAN X, WEI L, et al . Di-(2-ethylhexyl) phthalate inhibits testosterone level through disturbed hypothalamic-pituitary-testis axis and ERK-mediated 5α-Reductase 2[J ] . Sci Total Environ, 2016, 563/564: 566-575. DOI: 10.1016/j.scitotenv.2016.04.145.

YANG X, CHEN Y H, SONG W X, et al . Review of the role of ferroptosis in testicular function[J ] . Nutrients, 2022, 14(24): 5268. DOI: 10.3390/nu14245268.

MOJICA-VILLEGAS M A, IZQUIERDO-VEGA J A, CHAMORRO-CEVALLOS G, et al . Protective effect of resveratrol on biomarkers of oxidative stress induced by iron/ascorbate in mouse spermatozoa[J ] . Nutrients, 2014, 6(2): 489-503. DOI: 10.3390/nu6020489.

FENG Q W, LIU Y M, ZOU L P, et al . Fluorene-9-bisphenol exposure damages the testis in mice through a novel mechanism of ferroptosis[J ] . Food Chem Toxicol, 2024, 184: 114385. DOI: 10.1016/j.fct.2023.114385.

MIOTTO G, ROSSETTO M, DI PAOLO M L, et al . Insight into the mechanism of ferroptosis inhibition by ferrostatin-1[J ] . Redox Biol, 2020, 28: 101328. DOI: 10.1016/j.redox.2019.101328.

ZHANG D Y, WU X L, XUE X, et al . Ancient dormant virus remnant ERVW-1 drives ferroptosis via degradation of GPX4 and SLC3A2 in schizophrenia[J ] . Virol Sin, 2024, 39(1): 31-43. DOI: 10.1016/j.virs.2023.09.001..

XIAO J T, ZHANG S S, TU B J, et al . Arsenite induces ferroptosis in the neuronal cells via activation of ferritinophagy[J ] . Food Chem Toxicol, 2021, 151: 112114. DOI: 10.1016/j.fct.2021.112114..

SUN Y Y, SHEN J C, ZENG L, et al . Role of autophagy in di-2-ethylhexyl phthalate (DEHP)-induced apoptosis in mouse Leydig cells[J ] . Environ Pollut, 2018, 243(Pt A): 563-572. DOI: 10.1016/j.envpol.2018.08.089.

WANG J K, ZHAO T X, CHEN J D, et al . Multiple transcriptomic profiling: p53 signaling pathway is involved in DEHP-induced prepubertal testicular injury via promoting cell apoptosis and inhibiting cell proliferation of Leydig cells[J ] . J Hazard Mater, 2021, 406: 124316. DOI: 10.1016/j.jhazmat.2020.124316..

MARCHO C, OLUWAYIOSE O A, PILSNER J R. The preconception environment and sperm epigenetics[J]. Andrology, 2020, 8(4): 924-942. DOI: 10.1111/andr.12753.

PEREIRA S C, MARTINS A D, MONTEIRO M P, et al . Expression of obesity-related genes in human spermatozoa affects the outcomes of reproductive treatments[J ] . F S Sci, 2021, 2(2): 164-175. DOI: 10.1016/j.xfss.2021.03.004.

WU Y F, LI J C, LI C, et al . Fat mass and obesity-associated factor (FTO)-mediated N6-methyladenosine regulates spermatogenesis in an age-dependent manner[J ] . J Biol Chem, 2023, 299(6): 104783. DOI: 10.1016/j.jbc.2023.104783.

ZHOU S M, LI J Z, CHEN H Q, et al . FTO-Nrf2 axis regulates bisphenol F-induced leydig cell toxicity in an m6A-YTHDF2-dependent manner[J ] . Environ Pollut, 2023, 325: 121393. DOI: 10.1016/j.envpol.2023.121393.

YANG Y F, REN J J, ZHANG J F, et al . FTO ameliorates doxorubicin-induced cardiotoxicity by inhibiting ferroptosis via P53-P21/Nrf2 activation in a HuR-dependent m6A manner[J ] . Redox Biol, 2024, 70: 103067. DOI: 10.1016/j.redox.2024.103067..

MOU Y H, WANG J, WU J C, et al . Ferroptosis, a new form of cell death: opportunities and challenges in cancer[J ] . J Hematol Oncol, 2019, 12(1): 34. DOI: 10.1186/s13045-019-0720-y.

浏览量

120

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

富马酸二甲酯通过激活NRF2/SLC7A11轴抑制肺上皮细胞铁死亡并调控巨噬细胞极化缓解高原低氧急性肺损伤

染色质可及性重塑上调节律因子NPAS2表达促进胃癌铁死亡抵抗的作用研究

GenX通过System Xc⁻-Gpx4-Fsp1铁死亡轴诱导耳蜗毛细胞损伤

微小RNA-21-5p通过STAT3/P53/SLC7A11轴抑制铁死亡减轻高氧性急性肺损伤

PCBP1调控铁稳态抑制铁死亡对抗镉神经毒性的作用研究

DEHP通过抑制Fto表达诱导睾丸间质细胞铁死亡

DEHP induces ferroptosis in testicular interstitial cells by inhibiting Fto expression

DOI：10.16016/j.2097-0927.202404119

摘要

Abstract

关键词

Keywords

references