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两种睾丸体细胞系的基因表达谱对介导不同生殖毒性的4-硝基苯酚反应不同。

Gene expression profiles of two testicular somatic cell lines respond differently to 4-nitrophenol mediating vary reproductive toxicity.

作者信息

Wei Jinhuan, Lu Tingting, Dong Fulu, Zhang Chong, Zhang Yonghui

机构信息

Basic Medical Research Center, Nantong University School of Medicine, Nantong, 226001, Jiangsu Province, China.

Basic Medical Research Center, Nantong University School of Medicine, Nantong, 226001, Jiangsu Province, China.

出版信息

Toxicology. 2021 Nov;463:152991. doi: 10.1016/j.tox.2021.152991. Epub 2021 Oct 18.

Abstract

4-Nitrophenol (PNP) has been extensively used in manufacturing for several decades. Its toxic effects on the male reproductive system have been reported, but the underlying mechanisms remain unclear. In this study, we utilized two testicular somatic cell lines (TM3 and TM4 cells) to explore the possible toxic effects of PNP on the male reproductive system. The activity of the cells after exposure to different doses of PNP (0.01, 0.1, 1, 10 and 100 μM) was evaluated. PNP treatment at 10 μM significantly inhibited cell viability, and 10 μM PNP was thus selected for subsequent experiments. Although PNP (10 μM) inhibited cell proliferation, promoted cell apoptosis, and changed the cell cycle distribution and ultrastructure in both types of cells, these effects were more significant in the TM4 cells. In addition, an Agilent mouse mRNA array was used to identify the gene expression differences between the control and PNP (10 μM) exposed TM3 and TM4 cells. The microarray analysis identified 67 and 1372 differentially expressed genes mainly concentrated in endothelial cell morphogenesis and anatomical structure development in TM3 cells and associated with cardiovascular system development and circulatory system development in TM4 cells. Moreover, a pathway analysis revealed that PNP not only predominately affected meiotic recombination and meiosis in TM3 cells, but also influenced axon guidance and developmental biology in TM4 cells. These results suggest that TM3 and TM4 cells exhibit different responses to PNP, which might mediate different toxic mechanisms.

摘要

4-硝基苯酚(PNP)已在制造业中广泛使用了数十年。其对雄性生殖系统的毒性作用已有报道,但潜在机制仍不清楚。在本研究中,我们利用两种睾丸体细胞系(TM3和TM4细胞)来探究PNP对雄性生殖系统可能的毒性作用。评估了细胞在暴露于不同剂量的PNP(0.01、0.1、1、10和100μM)后的活性。10μM的PNP处理显著抑制了细胞活力,因此选择10μM的PNP用于后续实验。尽管PNP(10μM)抑制了两种细胞类型的细胞增殖、促进了细胞凋亡,并改变了细胞周期分布和超微结构,但这些作用在TM4细胞中更为显著。此外,使用安捷伦小鼠mRNA芯片来鉴定对照和暴露于PNP(10μM)的TM3和TM4细胞之间的基因表达差异。芯片分析确定了67个和1372个差异表达基因,主要集中在TM3细胞的内皮细胞形态发生和解剖结构发育以及与TM4细胞的心血管系统发育和循环系统发育相关。此外,通路分析表明,PNP不仅主要影响TM3细胞中的减数分裂重组和减数分裂,还影响TM4细胞中的轴突导向和发育生物学。这些结果表明,TM3和TM4细胞对PNP表现出不同的反应,这可能介导不同的毒性机制。

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