Plin4 exacerbates cadmium-decreased testosterone level via inducing ferroptosis in testicular Leydig cells
Xudong Zhang, Jian‐Jun Sun, Xinmei Zheng, Jin Zhang, Lulu Tan, Long-Long Fan, Ye‐Xin Luo, Y. Hu, Shendong Xu, Huan Zhou, Yufeng Zhang, Hao Li, Zhi Yuan, Wei Tian, Hua-Long Zhu, De‐Xiang Xu, Yong-Wei Xiong, Hua Wang
Abstract
Strong evidence indicates that environmental stressors are the risk factors for male testosterone deficiency (TD). However, the mechanisms of environmental stress-induced TD remain unclear. Based on our all-cause male reproductive cohort, we found that serum ferrous iron (Fe 2 ⁺) levels were elevated in TD donors. Then, we explored the role and mechanism of ferroptosis in environmental stress-reduced testosterone levels through in vivo and in vitro models. Data demonstrated that ferroptosis and lipid droplet deposition were observed in environmental stress-exposed testicular Leydig cells. Pretreatment with ferrostatin-1 (Fer-1), a specific ferroptosis inhibitor, markedly mitigated environmental stress-reduced testosterone levels. Through screening of core genes involved in lipid droplets formation, it was found that environmental stress significantly increased the levels of perilipins 4 (PLIN4) protein and mRNA in testicular Leydig cells. Further experiments showed that Plin4 siRNA reversed environmental stress-induced lipid droplet deposition and ferroptosis in Leydig cells. Additionally, environmental stress increased the levels of METTL3, METTL14, and total RNA m6A in testicular Leydig cells. Mechanistically, S-adenosylhomocysteine, an inhibitor of METTL3 and METTL14 heterodimer activity, restored the abnormal levels of Plin4 , Fe 2 ⁺ and testosterone in environmental stress-treated Leydig cells. Collectively, these results suggest that Plin4 exacerbates environmental stress-decreased testosterone level via inducing ferroptosis in testicular Leydig cells. • Human serum ferrous iron levels are negatively correlated with testosterone levels. • Environmental stress reduces testosterone levels by triggering ferroptosis in mouse testicular Leydig cells. • Environmental stress induces ferroptosis via enhancing Plin4 -mediated lipid droplet deposition in testicular Leydig cells. • Environmental stress drives m6A modification to upregulate Plin4 in mouse testicular Leydig cells.