Li Hong-Mei, Gao Yan-Rong, Liu Chang, Sheng Yu-Xin, Pu Ya-Jia, Sun Jia-He, Tian Ya-Nan, Yang Li, Ma Hui-Ming, Xu Hai-Ming
The Key Laboratory of Fertility Preservation and Maintenance of the Ministry of Education, Ningxia Medical University, Yinchuan 750004, China.
School of Basic Medicine, Ningxia Medical University, Yinchuan 750004, China.
Int J Mol Sci. 2024 Dec 15;25(24):13434. doi: 10.3390/ijms252413434.
The male reproductive impairment caused by environmental estrogens (EEs) stands as a pivotal research area in environmental toxicology. Alpha2-macroglobulin (A2M) emerges as a promising molecule capable of counteracting oxidative stress induced by EEs. This study conducted exposure experiments spanning PND1 to PND56 employing ICR mice, aiming to delve into the expression patterns of A2M and its modulated IL-6 in the testicular tissue of mice subsequent to diethylstilbestrol (DES) and benzophenone (BP) exposure, while elucidating the pivotal role of ERs in this intricate process. Our findings revealed that upon DES exposure (10 and 100 nM), there was a pronounced upregulation of A2M (mRNA and in situ protein levels) in mouse testicular tissue. Similarly, exposure to BPs (BP-1, BP-2, and BP-3, each at 10 and 1000 nM) exhibited comparable effects and increasing A2M levels in serum. Notably, BP exposure also caused an elevation in IL-6 levels (which could be directly regulated by A2M) within testicular tissue (mRNA and in situ protein). Remarkably, the specific estrogen receptor antagonist ICI 182780 (0.5 mg/kg/day) was effective in reversing the upregulation of both A2M and IL-6 induced by BP exposure. Significantly, the results of theoretical prediction of the potential ERE site in the A2m gene promoter region and ChIP-qPCR experiment provide essential and strong evidence for the key conclusion that A2m is the target gene of ER. Taken together, our study highlights EEs' ability to regulate A2M expression in the male reproductive system via the ER signaling pathway. This vital insight deepens our understanding of molecular mechanisms protecting against oxidative stress caused by EEs.
环境雌激素(EEs)导致的雄性生殖损伤是环境毒理学中的一个关键研究领域。α2-巨球蛋白(A2M)是一种有潜力的分子,能够对抗EEs诱导的氧化应激。本研究使用ICR小鼠进行了从出生后第1天(PND1)到第56天(PND56)的暴露实验,旨在探究在己烯雌酚(DES)和二苯甲酮(BP)暴露后,小鼠睾丸组织中A2M及其调控的白细胞介素-6(IL-6)的表达模式,同时阐明雌激素受体(ERs)在这一复杂过程中的关键作用。我们的研究结果显示,在DES暴露(10和100 nM)后,小鼠睾丸组织中A2M(mRNA和原位蛋白水平)显著上调。同样,暴露于BPs(BP-1、BP-2和BP-3,均为10和1000 nM)也表现出类似的效果,并使血清中A2M水平升高。值得注意的是,BP暴露还导致睾丸组织中IL-6水平(可由A2M直接调控)升高(mRNA和原位蛋白水平)。显著的是,特异性雌激素受体拮抗剂ICI 182780(0.5 mg/kg/天)能够有效逆转BP暴露诱导的A2M和IL-6上调。重要的是,A2m基因启动子区域潜在雌激素反应元件(ERE)位点的理论预测结果以及染色质免疫沉淀定量PCR(ChIP-qPCR)实验结果为“A2m是ER的靶基因”这一关键结论提供了重要且有力的证据。综上所述,我们的研究突出了EEs通过ER信号通路调节雄性生殖系统中A2M表达的能力。这一重要见解加深了我们对抵御EEs引起的氧化应激分子机制的理解。