College of Life Sciences, Key Laboratory of Animal Reproduction and Germplasm Enhancement in Universities of Shandong, Qingdao Agricultural University, Qingdao 266109, China.
Theranostics. 2021 Mar 5;11(10):4992-5009. doi: 10.7150/thno.55006. eCollection 2021.
Accumulated evidence indicates that environmental plasticizers are a threat to human and animal fertility. Di (2-ethylhexyl) phthalate (DEHP), a plasticizer to which humans are exposed daily, can trigger reproductive toxicity by acting as an endocrine-disrupting chemical. In mammals, the female primordial follicle pool forms the lifetime available ovarian reserve, which does not undergo regeneration once it is established during the fetal and neonatal period. It is therefore critical to examine the toxicity of DEHP regarding the establishment of the ovarian reserve as it has not been well investigated. The ovarian cells of postnatal pups, following maternal DEHP exposure, were prepared for single cell-RNA sequencing, and the effects of DEHP on primordial follicle formation were revealed using gene differential expression analysis and single-cell developmental trajectory. In addition, further biochemical experiments, including immunohistochemical staining, apoptosis detection, and Western blotting, were performed to verify the dataset results. Using single-cell RNA sequencing, we revealed the gene expression dynamics of female germ cells and granulosa cells following exposure to DEHP in mice. Regarding germ cells: DEHP impeded the progression of follicle assembly and interfered with their developmental status, while key genes such as , and others, strongly evidenced the reduction. As for granulosa cells: DEHP likely inhibited their proliferative activity, and activated the regulation of cell death. Furthermore, the interaction between ovarian cells mediated by transforming growth factor-beta signaling, was disrupted by DEHP exposure, since the expression of GDF9, BMPR1A, and SMAD3 was affected. In addition, DNA damage and apoptosis were elevated in germ cells and/or somatic cells. These findings offer substantial novel insights into the reproductive toxicity of DEHP exposure during murine germ cell cyst breakdown and primordial follicle formation. These results may enhance the understanding of DEHP exposure on reproductive health.
越来越多的证据表明，环境中的塑化剂对人类和动物的生育能力构成威胁。邻苯二甲酸二（2-乙基己基）酯（DEHP）是一种人类日常接触的塑化剂，它可以作为一种内分泌干扰化学物质引发生殖毒性。在哺乳动物中，雌性原始卵泡池形成一生中可用的卵巢储备，一旦在胎儿和新生儿期建立，就不会再生。因此，检查 DEHP 对卵巢储备建立的毒性至关重要，因为它尚未得到充分研究。 经母体 DEHP 暴露后的新生仔猪卵巢细胞进行单细胞 RNA 测序，通过基因差异表达分析和单细胞发育轨迹揭示 DEHP 对原始卵泡形成的影响。此外，还进行了进一步的生化实验，包括免疫组织化学染色、凋亡检测和 Western blot，以验证数据集结果。 通过单细胞 RNA 测序，我们揭示了暴露于 DEHP 后雌性生殖细胞和颗粒细胞的基因表达动态。对于生殖细胞：DEHP 阻碍了卵泡组装的进展并干扰了它们的发育状态，而 和其他关键基因则强烈表明其减少。对于颗粒细胞：DEHP 可能抑制了它们的增殖活性，并激活了细胞死亡的调节。此外，卵巢细胞之间的相互作用被 DEHP 暴露所破坏，因为 GDF9、BMPR1A 和 SMAD3 的表达受到影响。此外，生殖细胞和/或体细胞中的 DNA 损伤和凋亡增加。 这些发现为 DEHP 暴露在小鼠生殖细胞囊泡破裂和原始卵泡形成过程中的生殖毒性提供了新的重要见解。这些结果可能增强对 DEHP 暴露对生殖健康影响的理解。
