DEHP-mediated oxidative stress leads to impaired testosterone synthesis in Leydig cells through the cAMP/PKA/SF-1/StAR pathway

Luchen Yang; Shengzhuo Liu; Pan Song; Zhenghuan Liu; Zhufeng Peng; Depei Kong; Jing Zhou; Xin Yan; Kai Ma; Yunfei Yu; Xiaoyang Liu; Qiang Dong

doi:10.1016/j.envpol.2024.125503

DEHP-mediated oxidative stress leads to impaired testosterone synthesis in Leydig cells through the cAMP/PKA/SF-1/StAR pathway

Environ Pollut. 2024 Dec 8:125503. doi: 10.1016/j.envpol.2024.125503. Online ahead of print.

Authors

Luchen Yang¹, Shengzhuo Liu¹, Pan Song¹, Zhenghuan Liu¹, Zhufeng Peng¹, Depei Kong¹, Jing Zhou¹, Xin Yan¹, Kai Ma¹, Yunfei Yu¹, Xiaoyang Liu¹, Qiang Dong¹

Affiliation

¹ Department of Urology, Institute of Urology, West China Hospital, Sichuan University, Chengdu 610041, Sichuan, PR China.

PMID: 39657860
DOI: 10.1016/j.envpol.2024.125503

Abstract

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 H₂O₂ 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.

Keywords: Di-2-ethylhexyl phthalate; Leydig cells; Mono-2-ethylhexyl ester; Oxidative stress; Testosterone synthesis.