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1.550025 贵阳,贵州医科大学公共卫生与健康学院卫生毒理学系教研室
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
[曹佳,E-mail: caojia1962@126.comCAO Jia, E-mail: caojia1962@126.com
[张爱华,E-mail: aihuagzykd@163.comZHANG Aihua, E-mail: aihuagzykd@163.com
收稿:2022-12-01,
修回:2023-02-28,
纸质出版:2023-05-15
移动端阅览
卢春升, 梁月辉, 韩飞, 等. 聚苯乙烯微塑料对睾丸间质细胞凋亡的影响[J]. 陆军军医大学学报, 2023,45(9):885-892.
Chunsheng LU, Yuehui LIANG, Fei HAN, et al. Effect of polystyrene microplastics on apoptosis of male germ cells[J]. Journal of Army Medical University, 2023, 45(9): 885-892.
卢春升, 梁月辉, 韩飞, 等. 聚苯乙烯微塑料对睾丸间质细胞凋亡的影响[J]. 陆军军医大学学报, 2023,45(9):885-892. DOI: 10.16016/j.2097-0927.202212007.
Chunsheng LU, Yuehui LIANG, Fei HAN, et al. Effect of polystyrene microplastics on apoptosis of male germ cells[J]. Journal of Army Medical University, 2023, 45(9): 885-892. DOI: 10.16016/j.2097-0927.202212007.
目的
2
探讨聚苯乙烯微塑料(polystyrene microplastics,PS-MPs)对睾丸间质细胞凋亡的影响及可能机制。
方法
2
通过比较毒理基因组学数据库(CTD)检索Polystyrene导致男性泌尿生殖系统疾病的相关基因,采用GO和KEGG富集分析涉及的功能及信号通路。使用小鼠睾丸间质细胞(TM3细胞)作为研究模型,采用粒径为5 μm和80 nm的PS-MPs(0、10、20、40 μg/mL)处理TM3细胞48 h,观察细胞形态,利用CCK-8法检测细胞活性,流式细胞术检测细胞凋亡,RT-qPCR和Western blot检测富集的信号通路中关键分子mRNA和蛋白水平表达变化。
结果
2
对CTD数据库筛选的基因进行GO和KEGG富集分析发现,PS-MP暴露相关变化基因明显富集到细胞凋亡功能和p53信号通路(
P
<0.05)。5 μm和80 nm PS-MPs暴露TM3细胞48 h后均能明显抑制细胞生长,诱导细胞凋亡显著增加(
P
<0.05);5 μm和80 nm PS-MPs暴露可上调p53信号通路中p53、Bax的mRNA和蛋白表达水平(
P
<0.05),上调下游靶基因Caspase3和Caspase9表达(
P
<0.05),抑制抗凋亡分子Bcl2表达(
P
<0.05)。
结论
2
PS-MPs暴露可能通过p53信号通路诱导睾丸间质TM3细胞的凋亡。
Objective
2
To investigate the effect of polystyrene microplastic (PS-MPs) on the apoptosis of male germ cells.
Methods
2
Genes related to male urogenital disorders caused by polystyrene were searched in the Comparative Toxicogenomics Database (CTD). Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used to analyze the involved functions and signaling pathways. A cell model was established based on mouse mesenchymal cells (TM3 cells)
and then 0
10
20 and 40 μg/mL PS-MPs with a particle size of 5 μm and 80 nm respectively were used to treat TM3 cells respectively for 48 h. Cell morphology was observed
cell viability was detected with CCK-8 assay
apoptosis by flow cytometry
and mRNA and protein expression levels of key molecules in the p53 signaling pathway were detected by RT-qPCR and Western blotting
respectively.
Results
2
GO and KEGG enrichment analyses showed that the genes screened from CTD database were found to be significantly enriched in cell apoptosis and p53 signaling pathway (
P
<
0.05). PS-MPs of 5 μm and 80 nm in size treatment for 48 h resulted
in significant inhibition of cell growth while enhancement in cell apoptosis (
P
<
0.05)
up-regulated the mRNA and protein levels of p53 and Bax in the p53 signaling pathway (
P
<
0.05)
enhanced downstream molecules Caspase3 and Caspase9 (
P
<
0.05)
and inhibited Bcl2 at mRNA and protein levels in TM3 cells (
P
<
0.05).
Conclusion
2
PS-MPs exposure may induce apoptosis of testicular mesenchymal TM3 cells
via
the p53 signaling pathway.
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