Hydrogen sulfide (H2S), a gaseous intracellular signal transducer, participates in multiple physiological and pathological conditions, including reproductive conditions, and disrupts spermatogenesis. The blood-testis barrier (BTB) plays a vital role in spermatogenesis. However, the effect of H₂S on the BTB and the underlying mechanism remain unclear. Herein, we examined the effect of H₂S and omega-3 polyunsaturated fatty acids (ω-3 PUFAs) on the BTB and testicular functions. ICR male mice were randomly divided into the following groups: control, H₂S exposure, and H₂S exposure with ω-3 PUFAs intervention. The sperm parameters (sperm concentration and sperm motility) declined in the H₂S group and improved in the ω-3 intervention group. BTB integrity was severely disrupted by H₂S, and the BTB-related gene levels (ZO-1, Occludin, Claudin 11) decreased; ω-3 supplementation could alleviate BTB disruption by upregulating BTB-related genes, and TM4 Sertoli cells had a similar trend in vitro. p38 MAPK phosphorylation was upregulated in the Na₂S treatment group and downregulated after ω-3 cotreatment. These findings suggest that H₂S can impair the BTB and that ω-3 PUFAs supplementation can attenuate H₂S toxicity in the male reproductive system. Our study elucidated the relationship between a gasotransmitter (H₂S) and the BTB and identified the potential therapeutic effect of ω-3 PUFAs.

硫化氢 (H2S) 是一种气态细胞内信号转换器，参与多种生理和病理条件，包括生殖条件，并破坏精子发生。血睾屏障 (BTB) 在精子发生中起着至关重要的作用。然而，H ₂ S 对BTB 的影响及其潜在机制仍不清楚。在此，我们研究了 H ₂ S 和 omega-3 多不饱和脂肪酸 (ω-3 PUFA) 对 BTB 和睾丸功能的影响。ICR 雄性小鼠随机分为以下组：对照组、H ₂ S 暴露组和具有ω-3 PUFAs 干预的H ₂ S 暴露组。精子参数（精子浓度和精子活力）在 H ₂S组并在ω-3干预组中有所改善。BTB完整性被H ₂ S严重破坏，BTB相关基因水平（ZO-1、Occludin、Claudin 11）降低；补充 ω-3 可以通过上调 BTB 相关基因来减轻 BTB 破坏，TM4 Sertoli 细胞在体外也有类似的趋势。p38 MAPK 磷酸化在 Na ₂ S 处理组中上调，在 ω-3 共处理后下调。这些发现表明 H ₂ S 可以损害 BTB，补充 ω-3 PUFA 可以减弱男性生殖系统中的H ₂ S 毒性。我们的研究阐明了气体发射器 (H ₂S) 和 BTB，并确定了 ω-3 PUFA 的潜在治疗效果。