目的 绘制低压低氧环境暴露下睾丸组织单细胞转录组图谱，并进行支持细胞多样性分析，以期为开展生殖毒性机制相关研究提供新思路。

方法 取健康雄性小鼠20只，采用随机数字表法分为对照组( n =10)和低压低氧组( n =10)。其中对照组于正常环境下饲养，低压低氧组暴露于低压缺氧环境(压强=14 kPa，氧含量=14.5%)。6周后，取两组小鼠睾丸组织，采用Singleron Matrix ^TM 单细胞平台与Illumina NovaSeq二代测序技术获取转录组图谱并进行聚类、拟时序轨迹分析、功能分析、转录因子和细胞通讯研究。

结果 成功构建睾丸组织单细胞转录组图谱，其包含6个样本，总计约49 027个细胞，覆盖11种细胞类型。通过非负矩阵分解算法可将支持细胞聚类为4个亚群。其中亚群3可能对缺氧刺激更为敏感，通过影响PTN表达水平、调控精母细胞的细胞周期，以减少低压低氧所引发的生殖细胞功能损伤，PTN-PTPR通路可能为支持细胞对精原细胞发挥调控作用的一个重要调控节点；亚群4可通过KITLG-KIT信号通路在低压低氧环境暴露后调节精原细胞的细胞周期，影响精子发育。

结论 基于单细胞测序技术首次揭示了支持细胞在低压低氧暴露中的分子基础和调控讯号，从单细胞层面深入探究了低压低氧环境刺激所引发的生殖毒性机制，为后续开展相关临床研究提供了新视角。

Objective To construct a single-cell transcriptomic map of testicular tissue under hypobaric hypoxia exposure and perform diversity analysis of supportive cells, aiming to provide new insights into the mechanisms of reproductive toxicity for future research.

Methods Twenty healthy male mice were randomly divided into a control group ( n =10) and a hypobaric hypoxia group ( n =10). The control group was raised under normal conditions, while the hypobaric hypoxia group was exposed to a low-pressure hypoxic environment(pressure=14 kPa, oxygen content=14.5%). After 6 weeks, testicular tissue from both groups of mice was collected, and the transcriptomic map was obtained using the Singleron Matrix ^TM single-cell platform and Illumina NovaSeq second-generation sequencing technology. Clustering, pseudo-temporal trajectory analysis, functional analysis, transcription factor and cellular communication research were conducted.

Results A single-cell transcriptomic map of testicular tissue was successfully constructed, including 6 samples with a total of approximately 49 027 cells covering 11 cell types. Supportive cells were clustered into 4 subgroups using non-negative matrix factorization algorithm. Subgroup 3 may be more sensitive to hypoxic stimulation and could reduce reproductive cell function damage caused by hypobaric hypoxia by affecting PTN expression levels and regulating the cell cycle of spermatogonia. The PTN - PTPR pathway may be an important regulatory node for supportive cells to regulate spermatogonia. Subgroup 4 could regulate the cell cycle of spermatogonia after exposure to hypobaric hypoxia through the KITLG - KIT signaling pathway, thus affecting sperm development.

Conclusions Based on single-cell sequencing technology, the molecular basis and regulatory signals of supportive cells under hypobaric hypoxia exposure is revealed for the first time. It provides an in-depth exploration of the mechanisms of reproductive toxicity induced by hypobaric hypoxia at the single-cell level, offering a new perspective for future clinical research in this field.