Hypoxia drives spermatogenic impairment by triggering lysosome-dependent death of Leydig cells via M1 macrophage polarization
10.16016/j.2097-0927.202510077
- VernacularTitle:缺氧通过驱动M1巨噬细胞极化诱发睾丸间质细胞溶酶体依赖性死亡导致生精功能受损
- Author:
Dan MENG
1
;
Wei HE
;
Jitao ZENG
Author Information
1. 陆军军医大学(第三军医大学)第一附属医院生殖医学中心
- Keywords:
hypoxia;
testis;
Leydig cells;
macrophages;
lysosome
- From:
Journal of Army Medical University
2025;47(23):2893-2902
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the critical role of hypoxia in impairing spermatogenesis via M1 macrophage polarization and subsequent induction of lysosome-dependent cell death(LDCD)in Leydig cells.Methods Ten eight-week-old male SD rats(weight 220～250 g)were randomly divided into normoxia(n=5)and hypoxia groups(n=5).Hypoxic models were established by housing rats in hypoxic chambers simulating 5 000 m altitude for 4 weeks.TM3 mouse Leydig cells underwent 1%O2 exposure for 2 d.A transwell co-culture system integrated RAW 264.7 macrophages and TM3 cells.Testicular histopathology was evaluated via HE staining(spermatogenic cells/tubular area).Testosterone levels were quantified by ELISA(BCA-quantified).Immunofluorescence assessed Leydig cells(CYP11A1+),M1 macrophages(CD68+/CD86+in tissue;iNOS+in cells),and lysosomal integrity(LysoTracker/CTSB staining).Single-cell RNA-seq data(PRJNA926018)with GSEA identified programmed cell death pathways.Results Hypoxia significantly reduced spermatogenic cell density(P<0.01),testosterone levels(P<0.01),and Leydig cell counts(P<0.01).scRNA-seq revealed decreased proportions of elongated spermatids,Leydig cells,and sertoli cells(P<0.01),but increased spermatocytes/spermatogonia(P<0.01),with significant LDCD enrichment in Leydig cells(P=0.002 7).Hypoxia induced testicular M1 macrophage infiltration(CD68+/CD86+)and elevated iNOS+in RAW 264.7 cells(P<0.01).Transwell co-culture showed hypoxia-treated macrophages,but not hypoxia alone,disrupted lysosomal membrane stability in TM3 cells.Conclusion Hypoxia impairs spermatogenesis by driving M1 macrophage polarization,triggering LDCD in Leydig cells,and reducing testosterone synthesis.
