College of Medicine and Health Sciences, China Three Gorges University,Yichang, Hubei 443002, China; Hubei Province Key Laboratory of Occupational Hazard Identification and Control, Wuhan University of Science and Technology, Wuhan 430065, China.
College of Basic Medical Sciences, China Three Gorges University, Yichang, Hubei 443002, China.
Ecotoxicol Environ Saf. 2024 Nov 15;287:117282. doi: 10.1016/j.ecoenv.2024.117282. Epub 2024 Nov 5.
Di-(2-ethylhexyl) phthalate (DEHP), a widely used plasticizer, could cause male reproductive toxicity by disrupting spermatogenesis. Piwi-interacting RNAs (piRNAs) are a small non-coding RNAs specifically highly expressed in the germline and interact with PIWI proteins to regulate spermatogenesis. Accumulating studies have confirmed that environmental poisons could induce male reproductive injury via altering piRNA expression. However, it remains unclear whether DEHP causes male reproductive dysfunction by perturbing piRNA expression levels. In this study, we conducted piRNA microarray expression analyses on testes of DEHP-exposed and control male rats and performed some in vitro and in vivo studies to explore the role of piRNA on DEHP-induced male reproductive toxicity. Our results showed that DEHP exposure leaded to changed expression profiles of piRNAs in pubertal male rat testes. And bioinformatics analyses revealed that down-regulated piR-rno-26751 probably targeted Insr mRNA expression regulation. Results from gene and protein expression tests demonstrated that DEHP caused decreased expression level of INSR mainly in spermatogonia. Moreover, MEHP, the main metabolite of DEHP resulted in cell apoptosis and down-regulation of INSR and its downstream p-IRS1, p-PI3K, p-AKT and p-FOXO1 in GC-1spg cells. Conversely, overexpression of INSR restored cell apoptosis and the down-regulation of the above proteins in GC-1spg cells. In conclusion, these findings suggest that DEHP-induced down-regulation of piR-rno-26751 targets the suppression of INSR, leading to apoptosis of spermatogonia in pubertal male rats.
邻苯二甲酸二（2-乙基己基）酯（DEHP）作为一种广泛使用的增塑剂，通过破坏精子发生导致雄性生殖毒性。Piwi 相互作用 RNA（piRNA）是一种在生殖细胞中特异性高度表达的小非编码 RNA，与 PIWI 蛋白相互作用以调节精子发生。越来越多的研究证实，环境毒物可以通过改变 piRNA 的表达来诱导雄性生殖损伤。然而，DEHP 是否通过干扰 piRNA 表达水平引起雄性生殖功能障碍仍不清楚。在这项研究中，我们对 DEHP 暴露和对照雄性大鼠的睾丸进行了 piRNA 微阵列表达分析，并进行了一些体内和体外研究，以探讨 piRNA 在 DEHP 诱导的雄性生殖毒性中的作用。我们的结果表明，DEHP 暴露导致青春期雄性大鼠睾丸中 piRNA 的表达谱发生变化。生物信息学分析显示，下调的 piR-rno-26751 可能靶向 Insr mRNA 表达调控。基因和蛋白表达测试的结果表明，DEHP 导致 INSR 的表达水平主要在精原细胞中降低。此外，DEHP 的主要代谢物 MEHP 导致 GC-1spg 细胞凋亡和 INSR 及其下游 p-IRS1、p-PI3K、p-AKT 和 p-FOXO1 的下调。相反，INSR 的过表达恢复了 GC-1spg 细胞中的细胞凋亡和上述蛋白的下调。总之，这些发现表明，DEHP 诱导的 piR-rno-26751 下调靶向抑制 INSR，导致青春期雄性大鼠精原细胞凋亡。
