Guo Li, Li Siqi, Li Zehui, Li Yuan, Li Jiashan, Cheng Huiting, Lu Yuan, Lin Na, Xu Ying
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16, Nanxiao Street, Dongzhimen Nei, Dongcheng District, Beijing 100700, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, No. 16, Nanxiao Street, Dongzhimen Nei, Dongcheng District, Beijing 100700, China.
Life Sci. 2025 Aug 26;380:123935. doi: 10.1016/j.lfs.2025.123935.
Tripterygium glycoside tablet (TGT), a first-line anti-inflammatory drug for rheumatoid arthritis (RA), is severely limited in clinical use by male reproductive toxicity with unclear mechanisms. This study aimed to confirm TGT's anti-RA efficacy, characterize its reproductive toxicity in collagen-induced arthritis (CIA) rats, and elucidate whether the NELL2-Lumicrine system mediates such damage.
TGT's anti-inflammatory efficacy in CIA rats was evaluated via arthritis scoring, histopathology, and cytokine quantification. Male reproductive toxicity was assessed using sperm analysis, fertilization evaluation, histopathology, and hormone measurement. NELL2-Lumicrine system targets were identified by immunofluorescence and Western blot. Integrated metabolomic and transcriptomic analyses explored TGT's effects on testicular metabolism and gene expression. Molecular docking, dynamics simulation, and cell co-culture experiments verified inhibitory effects of TGT's bioactive components on the NELL2-Lumicrine system.
TGT alleviated joint swelling, reduced arthritis scores, and decreased serum TNF-α/IL-1β in CIA rats. However, TGT impaired sperm quality and fertilization capacity, increased sperm/embryo abnormalities, and induced testicular-epididymal damage, with hypothalamic-pituitary-gonadal axis hormone dysregulation. Mechanistically, TGT downregulated NELL2 and downstream targets (ROS1, OVCH2, ADAM28, ADAM3), disrupting sperm maturation. Multi-omics revealed TGT disrupted testicular steroid biosynthesis and Fgf signaling linked to the NELL2-Lumicrine system. Computational modeling and in vitro experiments confirmed triptolide and triptonide bind NELL2 strongly, suppressing its signaling pathway.
Our data provide the first evidence that TGT induces male reproductive toxicity by inhibiting the NELL2-Lumicrine system, downregulating NELL2 and its downstream targets (ROS1, OVCH2, ADAM28, and ADAM3), which are essential for sperm maturation.
雷公藤多苷片（TGT）是类风湿性关节炎（RA）的一线抗炎药物，但由于其男性生殖毒性机制不明，临床应用受到严重限制。本研究旨在确认TGT的抗RA疗效，明确其在胶原诱导性关节炎（CIA）大鼠中的生殖毒性，并阐明NELL2-Lumicrine系统是否介导了这种损伤。
通过关节炎评分、组织病理学和细胞因子定量评估TGT在CIA大鼠中的抗炎疗效。使用精子分析、受精评估、组织病理学和激素测量来评估男性生殖毒性。通过免疫荧光和蛋白质印迹鉴定NELL2-Lumicrine系统靶点。综合代谢组学和转录组学分析探索TGT对睾丸代谢和基因表达的影响。分子对接、动力学模拟和细胞共培养实验验证了TGT生物活性成分对NELL2-Lumicrine系统的抑制作用。
TGT减轻了CIA大鼠的关节肿胀，降低了关节炎评分，并降低了血清TNF-α/IL-1β水平。然而，TGT损害了精子质量和受精能力，增加了精子/胚胎异常，并导致睾丸-附睾损伤，伴有下丘脑-垂体-性腺轴激素失调。机制上，TGT下调了NELL2及其下游靶点（ROS1、OVCH2、ADAM28、ADAM3），破坏了精子成熟。多组学研究表明，TGT破坏了与NELL2-Lumicrine系统相关的睾丸类固醇生物合成和Fgf信号通路。计算模型和体外实验证实，雷公藤甲素和雷公藤内酯酮与NELL2紧密结合，抑制其信号通路。
我们的数据首次证明，TGT通过抑制NELL2-Lumicrine系统，下调对精子成熟至关重要的NELL2及其下游靶点（ROS1、OVCH2、ADAM28和ADAM3），从而诱导男性生殖毒性。
