睾丸损伤和影响精子发生是甲氨蝶呤 (MTX) 使用的主要并发症。氧化应激是导致炎症和细胞凋亡诱导的过程之一。吡非尼酮 (PFD) 是一种著名的抗纤维化药物，用于治疗间质性肺纤维化，此外还具有抗炎、抗氧化和抗凋亡功能。该研究旨在探讨 PFD 在 MTX 诱导的睾丸病大鼠模型中提供的潜在保护作用。实验设计包括四组，每组包含七只成年 Wistar 大鼠：对照组、PFD（500 mg/kg/天，口服）、MTX（0.5 mg/kg，腹腔注射，每周两次）和 PFD/MTX 治疗组。治疗持续 4 周。采集血液和睾丸样本用于生化、组织学、免疫组织化学和聚合酶链反应（PCR）分析。此外，睾丸损伤和生精活性分别通过睾丸损伤和Johnsen评分系统进行分级。 PFD对血清睾酮（TST）水平产生积极影响，减少睾丸炎症介质[肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）]，降低睾丸氧化负担，增加超氧化物歧化酶（SOD） ，并保护睾丸组织学结构。此外，还记录了抗纤维化作用、抗 caspase-3 和 PCNA 增强活性。 PFD 表现出保护潜力，通过抑制睾丸氧化应激、炎症、纤维化和细胞凋亡来减轻 MTX 诱导的睾丸病变，并通过组织学、免疫组织化学和生化方法证实保留睾丸增殖功效。© 2024。作者，获得 Springer-Verlag GmbH 德国（Springer Nature 旗下公司）的独家许可。

Testicular injury and affected spermatogenesis are major complications of methotrexate (MTX) use. Oxidative stress is one contributing process leading to inflammation and apoptosis induction. Pirfenidone (PFD) is a well-known anti-fibrotic drug prescribed for interstitial lung fibrosis, in addition to anti-inflammatory, antioxidative, and antiapoptotic capabilities. The study aimed to explore the potential protection afforded by PFD in a rat model of MTX-induced testiculopathy. The experimental design included four groups, each containing seven adult Wistar rats: control, PFD (500 mg/kg/day, orally)-, MTX (0.5 mg/kg, intraperitoneal, twice weekly)-, and PFD/MTX-treated groups. Treatment continued for 4 weeks. Blood and testicular samples were harvested for biochemical, histological, immunohistochemical, and polymerase chain reaction (PCR) analyses. Also, the testicular damage and spermatogenic activity were graded by the testicular injury and Johnsen scoring system, respectively. PFD positively affected the serum testosterone (TST) level, reduced the testicular inflammatory mediators [tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β)], reduced the testicular oxidative burden, increased superoxide dismutase (SOD), and protected the testicular histological structure. In addition, antifibrotic effects, anti-caspase-3, and PCNA enhancement activity were recorded. PFD exhibited a protective potential and mitigated the MTX-induced testiculopathy via suppression of testicular oxidative stress, inflammation, fibrosis, and apoptosis and retaining the testicular proliferative efficacy as confirmed by histological, immunohistochemical, and biochemical methods.© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.