铁死亡参与热应激下小鼠生精障碍OA
Ferroptosis is involved in spermatogenic dysfunction in mice under heat stress
目的 探究铁死亡与热应激下小鼠生精障碍的相关性.方法 将成年雄性 C57BL/6 小鼠分为对照组(control)和热应激组(HS),收集附睾管中的精子,使用精子活力检测仪检测精子活力等指标;采集小鼠睾丸,利用RNA-Seq 技术进行差异基因及 GO 和 KEGG 分析;使用铁死亡抑制剂 Fer1 进行 rescue 实验,即增加 Fer1-HS 组,采用 HE 和 TUNEL 染色分别检测睾丸组织结构变化;利用试剂盒分别检测睾丸组织内铁死亡相关指标(Fe2+和 GSH)的改变.结果 HS 可导致小鼠精子活力和浓度有降低迹象,生精小管松散,生精上皮变薄,生精细胞大量凋亡;且睾丸组织 Fe2+水平显著高于对照组(P＜0.05),而 GSH 水平有降低的趋势.RNA-seq 测序筛选到了铁死亡相关基因 Miox、Prap1 和 Mt3;GO 分析发现热应激可使小鼠睾丸铁离子和脂质过氧化物等 GO 通路更活跃.而 Fer1 处理小鼠后,Fer1-HS 组睾丸生精小管松散程度和生精细胞脱落和HS 组相比均有所恢复;Fer1-HS 组小鼠睾丸Fe2+水平和 HS 组相比显著下降(P＜0.05),而 GSH 水平和 HS 组相比有回升迹象.结论 铁死亡参与诱导热应激下小鼠睾丸生精障碍,且主要表现为铁水平过高.
Objective To investigate the correlation between ferroptosis and spermatogenic dysfunction in mice un-der heat stress.Methods Adult male C57BL/6 mice were divided into the control group and the heat stress(HS)group.Spermatozoa were collected from the epididymal ducts,and indicators such as sperm motility were detected using a sperm motility analyzer.Mouse testes were collected,and RNA-Seq technology was used for differential gene analysis,as well as Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.A rescue experiment was conducted using the ferroptosis inhibitor Ferrostatin-1(Fer1),with an additional Fer1-HS group established.Hematoxylin-eosin(HE)staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining were used to detect changes in testicular tissue structure,respectively.Kits were used to detect changes in ferroptosis-related indicators(Fe2+and glutathione,GSH)in testicular tissue.Results Heat stress(HS)tended to reduce sperm motility and concentration in mice,caused loosening of seminiferous tubules,thinning of the spermatogenic epithelium,and massive apoptosis of spermatogenic cells.Moreover,the lev-el of Fe2+in testicular tissue was significantly higher than that in the control group(P＜0.05),while the GSH level showed a downward trend.RNA-seq identified ferroptosis-related genes Miox,Prap1,and Mt3.GO analysis re-vealed that heat stress enhanced the activity of GO pathways related to iron ions and lipid peroxides in mouse testes(Intracellular sequestering of iron ion,cellular lipid catabolic process and negative regulation of oxidoreductase ac-tivity).After treatment with Fer1,the Fer1-HS group showed partial recovery in the loosening degree of testicular seminiferous tubules and the shedding of spermatogenic cells compared with the HS group.Additionally,the Fe2+level in the testes of the Fer1-HS group was significantly lower than that in the HS group(P＜0.05),while the GSH level showed a tendency to increase compared with the HS group.Conclusions Ferroptosis is involved in inducing testicular spermatogenic dysfunction in mice under heat stress,mainly manifested by excessive iron levels.
徐子迅;陈相廷;孙骜;贾茗迪;夏青
上海交通大学医学院 基础医学实验教学中心,上海 200025上海交通大学医学院 基础医学实验教学中心,上海 200025上海交通大学医学院 基础医学实验教学中心,上海 200025上海交通大学医学院 基础医学实验教学中心,上海 200025上海交通大学医学院 基础医学实验教学中心,上海 200025
医药卫生
铁死亡热应激睾丸生精障碍
ferroptosisheat stresstestisspermatogenic dysfunction
《基础医学与临床》 2026 (10)
1316-1321,6
上海高校实验队伍建设计划(BJ1-3000-22-0085)
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