Effect of polystyrene micro/nanoplastics on mesenchymal phenotypic transformation in testicular Sertoli cells

CAI Jing; WEN Yixian; ZHANG Huilian; LI Yi; HAN Fei

doi:10.16016/j.2097-0927.202502043

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Effect of polystyrene micro/nanoplastics on mesenchymal phenotypic transformation in testicular Sertoli cells

Monographic Reports | 更新时间：2025-07-27

- Effect of polystyrene micro/nanoplastics on mesenchymal phenotypic transformation in testicular Sertoli cells
- Journal of Army Medical University Vol. 47, Issue 15, Pages: 1708-1719(2025)
- 作者机构：
  
  重庆医科大学公共卫生学院，生殖与发育教育部国际合作联合实验室，重庆
- 作者简介：
  
  HAN Fei， E-mail： han16897723@163.com
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(82073137)
- DOI：10.16016/j.2097-0927.202502043
  CLC： R339.21;R994.6;TQ325.2
- Received：14 February 2025，
  
  Revised：25 April 2025，
  
  Accepted：20 May 2025，
  
  Published：15 August 2025
- 稿件说明：
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蔡景，文乙先，张慧莲，等.聚苯乙烯微/纳米塑料对睾丸支持细胞间质表型转化的影响［J］.陆军军医大学学报，2025，47（15）：1708-1719.DOI：10.16016/j.2097-0927.202502043.

CAI J，WEN Y X，ZHANG H L，et al.Effect of polystyrene micro/nanoplastics on mesenchymal phenotypic transformation in testicular Sertoli cells［J］.J Army Med Univ，2025，47（15）：1708-1719.DOI：10.16016/j.2097-0927.202502043.
蔡景，文乙先，张慧莲，等.聚苯乙烯微/纳米塑料对睾丸支持细胞间质表型转化的影响［J］.陆军军医大学学报，2025，47（15）：1708-1719.DOI：10.16016/j.2097-0927.202502043. DOI：

CAI J，WEN Y X，ZHANG H L，et al.Effect of polystyrene micro/nanoplastics on mesenchymal phenotypic transformation in testicular Sertoli cells［J］.J Army Med Univ，2025，47（15）：1708-1719.DOI：10.16016/j.2097-0927.202502043. DOI：

摘要

目的

探讨聚苯乙烯微/纳米塑料（polystyrene micro/nanoplastics，PS-MNPs）对睾丸支持细胞的影响及潜在机制。

方法

60只8周龄C57BL/6N雄性小鼠按照完全随机法分为（

=20）：Ctrl组（去离子水）、聚苯乙烯纳米塑料（polystyrene nanoplastics， PS-NPs）组［粒径为20 nm， 2.5 mg/（kg·d）］和聚苯乙烯微米塑料（polystyrene microplastics， PS-MPs）组［粒径为5 μm， 2.5 mg/（kg·d）］。每日灌胃1次，持续6个月。采用HE染色分析睾丸组织病理形态差异；标志蛋白免疫组化分析支持细胞数量变化。采用GO和KEGG富集分析睾丸转录组富集的功能及信号通路。使用小鼠睾丸支持细胞系（TM4）作为研究模型，分为Con组（去离子水）、2.5NPs组（粒径为20 nm， 2.5 μg/mL）和2.5MPs组（粒径为5 μm， 2.5 μg/mL），各组TM4细胞均连续处理至130代，CCK-8法及EdU检测细胞活性与增殖能力，Transwell和细胞划痕实验检测细胞迁移能力，利用RT-qPCR和Western blot检测调控间质表型转化关键分子mRNA和蛋白水平表达变化。

结果

病理学分析结果显示，与Ctrl组相比，PS-NPs和PS-MPs组小鼠的睾丸曲细精管间距增大，生精细胞排列松散，并伴有空泡化程度增加。标志蛋白免疫组化结果显示，PS-NPs和PS-MPs组小鼠的睾丸支持细胞数量与Ctrl组相比呈下降趋势，其中PS-NPs组差异具有统计学意义（

0.01）。对暴露睾丸组织转录组筛选的差异基因进行GO和KEGG富集分析发现，PS-MNPs暴露相关变化基因明显富集到细胞黏附分子信号通路（

0.05）。PS-MPs暴露TM4细胞明显抑制细胞生长及细胞迁移能力（

0.05）；PS-NPs暴露TM4细胞对细胞生长没有明显影响，但能明显增强细胞迁移能力；PS-NPs暴露后抑制E-cadherin和ZO-1的蛋白表达水平（

0.01），上调N-cadherin和Vimentin的表达（

0.01）；PS-MPs暴露后上调Vimentin的表达（

0.01），下调E-cadherin、N-cadherin及ZO-1的表达（

0.05）；PS-MPs和PS-NPs暴露上调Snail2、Twist1和Zeb2的mRNA水平（

0.01）。

结论

PS-MNPs暴露会导致TM4细胞增殖和迁移功能异常，并可能通过间质表型转化诱导支持细胞与各级生精细胞间的细胞-细胞接触减少，进而导致睾丸损伤。

Abstract

Objective‍　‍To investigate the effects and potential mechanisms of polystyrene micro/nanoplastics (PS-MNPs) on testicular Sertoli cells. Methods Sixty male C57BL/6N mice (8 weeks old) were randomly divided into a control group (deionized water)

a PS-NPs group [particle size of 20 nm

2.5 mg/(kg·d)

]

and a PS-MPs group [particle size of 5 μm

2.5 mg/(kg·d)

]

with 20 mice in each group. The corresponding agents were gavaged once daily for 6 months. HE staining was used to observe the histopathological and morphological changes in the testicular tissues. Immunohistochemistry of marker proteins was employed to evaluate the changes in the number of Sertoli cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to identify functions and signaling pathways enriched in the testicular transcriptome. Mouse testicular Sertoli cell line TM4 was divided into a control group (deionized water)

a 2.5NPs group (2.5 μg/mL)

and a 2.5MPs group (2.5 μg/mL). All groups received continuous exposure through 130 cell passages. Cell viability and proliferative capacity were evaluated using CCK-8 assay and EdU incorporation

while cell migration was assessed using transwell and cell scratch assays. RT-qPCR and Western blotting were used to detect the changes in the expression of key molecules regulating mesenchymal phenotypic transformation (MPT) at mRNA and protein levels. Results Pathological analysis revealed that

when compared to the control group

PS-NPs and PS-MPs treatment resulted in extended spaces between testicular seminiferous tubules

loosely arranged spermatogenic cells

and enhanced vacuolization. Immunohistochemical analysis of marker proteins indicated a decreasing trend in the number of testicular Sertoli cells in the PS-NPs and PS-MPs groups than the control group

with the PS-NPs group having stat

istical significance (

0.01). GO and KEGG enrichment analyses revealed that PS-MNPs exposure-related altered genes were significantly enriched in cell adhesion signaling pathways (

0.05). PS-MPs exposure significantly inhibited the growth and migration ability of TM4 cells (

0.05)

but PS-NPs exposure had no such effect on cell growth but notably enhanced cell migration ability. PS-NPs exposure inhibited the expression levels of E-cadherin and ZO-1 (

0.01) and up-regulated the expression of N-cadherin and vimentin (

0.01)

and PS-MPs exposure led to significant up-regulation of vimentin (

0.01) and down-regulation of E-cadherin

N-cadherin

and ZO-1 (

0.05). Both PS-MPs and PS-NPs exposure up-regulated the mRNA levels of Snail2

Twist1

and Zeb2 (

0.01). Conclusion Exposure of PS-MNPs leads to abnormal proliferation and migration of TM4 cells

induces decreases in cell-cell contacts among Sertoli cells and spermatogenic cells at all levels possibly through MPT

and thus results in testicular damage.

关键词

Keywords

references

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