Lin Hang, Su Ming, Zhu He, Yu Yang, Sang Jianmin, Wang Yiyan, Zhu Qiqi, Zhu Yang, Li Xiaoheng, Li Xingwang, Ge Ren-Shan, Li Huitao
Department of Anesthesiology and Perioperative Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Key Laboratory of Pediatric Anesthesiology, Ministry of Education, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Key Laboratory of Anesthesiology of Zhejiang Province, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Key Laboratory of Environment and Male Reproductive Medicine of Wenzhou, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
Department of Anesthesiology and Perioperative Medicine, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Key Laboratory of Pediatric Anesthesiology, Ministry of Education, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China; Key Laboratory of Anesthesiology of Zhejiang Province, Wenzhou Medical University, Wenzhou, Zhejiang 325000, China.
Ecotoxicol Environ Saf. 2025 Jan 1;289:117503. doi: 10.1016/j.ecoenv.2024.117503. Epub 2024 Dec 12.
This research investigated the impact of cyclopiazonic acid (CPA), a mycotoxin, on the function of progenitor Leydig cells (PLCs) in prepubertal male rats, focusing on its potential disruption of mitochondrial integrity through mitofusin 1 (MFN1) modulation. In vivo, Sprague Dawley rats received CPA (0.2, 1, 5 mg/kg/day) via gavage from postnatal days 21-28 to evaluate PLC function and mitochondrial morphology using serum hormone levels, histology, qPCR, and Western blot analyses. In vitro, rat R2C cells were treated with CPA (0.1, 1, 10 μM) alone or in combination with 100 μM leflunomide to assess PLC development through testosterone measurements, Western blotting, flow cytometry, and Mito-Tracker Green Staining. The findings from in vivo experiments showed that CPA reduced serum testosterone and progesterone levels at 1 mg/kg/day. The qPCR and Western blotting analyses revealed significant alterations in the expression of genes and proteins pertinent to PLC function, such as Scarb1, Star, Cyp11a1, and Cyp17a1. Immunofluorescence staining further revealed a reduction in MFN1 expression following exposure to CPA. In vitro experiments corroborated these observations, demonstrating that CPA induced mitochondrial fragmentation by downregulating SIRT1, PGC1-α, MFN1, and OPA1, increase reactive oxygen species, and inhibit testosterone synthesis in R2C cells. The administration of leflunomide was shown to mitigate the detrimental effects of CPA on PLCs. In conclusion, this research sheds new light on the deleterious effects of CPA on the reproductive development of prepubertal males.
本研究调查了霉菌毒素环匹阿尼酸（CPA）对青春期前雄性大鼠睾丸间质祖细胞（PLCs）功能的影响，重点关注其通过调节线粒体融合蛋白1（MFN1）对线粒体完整性的潜在破坏作用。在体内实验中，从出生后第21天至28天，给斯普拉格-道利大鼠经口灌胃给予CPA（0.2、1、5 mg/kg/天），通过血清激素水平、组织学、qPCR和蛋白质印迹分析来评估PLC功能和线粒体形态。在体外实验中，将大鼠R2C细胞单独用CPA（0.1、1、10 μM）处理，或与100 μM来氟米特联合处理，通过睾酮测定、蛋白质印迹、流式细胞术和线粒体追踪绿染色来评估PLC的发育。体内实验结果表明，CPA在剂量为1 mg/kg/天时降低了血清睾酮和孕酮水平。qPCR和蛋白质印迹分析显示，与PLC功能相关的基因和蛋白质表达发生了显著变化，如Scarb1、Star、Cyp11a1和Cyp17a1。免疫荧光染色进一步显示，暴露于CPA后MFN1表达降低。体外实验证实了这些观察结果，表明CPA通过下调SIRT1、PGC1-α、MFN1和OPA1诱导线粒体碎片化，增加活性氧，并抑制R2C细胞中的睾酮合成。来氟米特的给药显示可减轻CPA对PLCs的有害影响。总之，本研究为CPA对青春期前男性生殖发育的有害影响提供了新的见解。
