Yang Luchen, Liu Shengzhuo, Song Pan, Liu Zhenghuan, Peng Zhufeng, Kong Depei, Zhou Jing, Yan Xin, Ma Kai, Yu Yunfei, Liu Xiaoyang, Dong Qiang
Department of Urology, Institute of Urology, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, PR China.
Department of Urology, Institute of Urology, West China Hospital, Sichuan University, Chengdu, 610041, Sichuan, PR China.
Environ Pollut. 2025 Feb 1;366:125503. doi: 10.1016/j.envpol.2024.125503. Epub 2024 Dec 8.
Leydig cells (LCs) injury is often irreversible upon discovery; hence, early identification of risk factors for injury is crucial. The ubiquitous plasticizer di-2-ethylhexyl phthalate (DEHP) in the environment has been shown to potentially cause damage to LCs. However, the underlying mechanisms remain unclear. The present study utilized scRNA-seq analysis, the advantage of which is the ability to explore the characteristics of various testicular cells, combined with studies in vitro and in vivo, to assay the changes in and damage processes of LCs during DEHP exposure. We found that DEHP disrupted the structure and function of LCs. GO analysis suggested that a series of pathways changed, among which the most significant were the "steroid synthesis" and "oxidative stress" pathways. Moreover, DEHP dramatically changed the manner of interaction between LCs and other cells, and the most significant type was the cell-cell contact, which included NECTIN, APP, CADM, and CD39. In addition, the activity of multiple transcription factors (TFs) decreased after DEHP exposure, and the activity of steroidogenic factor 1 (SF-1, Nr5a1) was the most obviously altered. Next, we found that the LCs region indeed experienced oxidative stress, including increased ROS signals, the decreased SOD activity and T-AOC, and increased concentration of 8-OHdG and MDA content. The testosterone level, as well as the expression of StAR, P450scc, and 3β-HSD, was also reduced. To study the association between testosterone synthesis and oxidative stress, the antioxidants N-acetyl-L-cysteine (NAC) and HO were used, and we found that mono-2-ethylhexyl ester (MEHP, a major biometabolite of DEHP) disrupted testosterone synthesis through the inhibition of the cAMP/PKA/SF-1/StAR pathway by inducing oxidative stress. Our study provides new insights into the role and mechanisms of DEHP in LCs injury.
睾丸间质细胞（LCs）损伤一旦被发现往往是不可逆的；因此，早期识别损伤的风险因素至关重要。环境中普遍存在的增塑剂邻苯二甲酸二（2-乙基己基）酯（DEHP）已被证明可能对睾丸间质细胞造成损害。然而，其潜在机制仍不清楚。本研究利用单细胞RNA测序（scRNA-seq）分析（其优势在于能够探究各种睾丸细胞的特征），并结合体外和体内研究，来检测DEHP暴露期间睾丸间质细胞的变化和损伤过程。我们发现DEHP破坏了睾丸间质细胞的结构和功能。基因本体（GO）分析表明一系列通路发生了变化，其中最显著的是“类固醇合成”和“氧化应激”通路。此外，DEHP显著改变了睾丸间质细胞与其他细胞之间的相互作用方式，最显著的类型是细胞间接触，其中包括NECTIN、APP、CADM和CD39。另外，DEHP暴露后多种转录因子（TFs）的活性降低，其中类固醇生成因子1（SF-1，Nr5a1）的活性改变最为明显。接下来，我们发现睾丸间质细胞区域确实经历了氧化应激，包括活性氧（ROS）信号增加、超氧化物歧化酶（SOD）活性和总抗氧化能力（T-AOC）降低，以及8-羟基脱氧鸟苷（8-OHdG）浓度和丙二醛（MDA）含量增加。睾酮水平以及类固醇急性调节蛋白（StAR）、细胞色素P450侧链裂解酶（P450scc）和3β-羟基类固醇脱氢酶（3β-HSD）的表达也降低。为了研究睾酮合成与氧化应激之间的关联，我们使用了抗氧化剂N-乙酰-L-半胱氨酸（NAC）和HO，并且我们发现单-2-乙基己基酯（MEHP，DEHP的一种主要生物代谢物）通过诱导氧化应激抑制环磷酸腺苷/蛋白激酶A/类固醇生成因子1/类固醇急性调节蛋白（cAMP/PKA/SF-1/StAR）通路来破坏睾酮合成。我们的研究为DEHP在睾丸间质细胞损伤中的作用和机制提供了新的见解。
