Jiangxi Provincial Key Laboratory for Animal Health, Institute of Animal Population Health, College of Animal Science and Technology, Jiangxi Agricultural University, No. 1101 Zhimin Avenue, Economic and Technological Development District, Nanchang 330045, Jiangxi, PR China.
Nanchang Institute of Technology Medical College, No. 901, Hero Avenue, Nanchang Economic Development Zone, Nanchang 330044, Jiangxi, PR China.
Phytomedicine. 2024 Dec;135:156245. doi: 10.1016/j.phymed.2024.156245. Epub 2024 Nov 9.
Male infertility is a worldwide concern that is associated with a decline in sperm quality. Environmental pollutants such as deltamethrin (DM) have harmful effects on male reproductive organs. By maintaining intracellular redox homeostasis, ginkgo biloba extract (GBE) can alleviate male reproductive dysfunction. However, research on the mechanisms by which GBE alleviates reproductive toxicity induced by DM is limited.
In this study, we investigated whether GBE can alleviate DM-induced testicular and Sertoli cell reproductive toxicity by modulating SKP2 and Beclin1, thus providing a theoretical basis for the development of novel therapeutic approaches.
We explored the role of GBE in mitigating DM-induced testicular damage, with a specific focus on the intricate involvement of ubiquitination and autophagy.
An experimental model was constructed using ICR male mice and the TM4 cell line. Tissue, cellular, and sperm morphological changes were observed through methods such as Hematoxylin and Eosin (H&E) staining, Periodate-Schiff (PAS) staining, ultrastructural observation, immunohistochemistry, and immunofluorescence. Enzyme and hormone levels were measured, and gene and protein levels were detected using real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting techniques.
In vivo experiments showed that DM exposure led to decreased sex hormone levels, increased seminiferous tubule diameter and impaired spermatogenesis. Meanwhile, DM exposure was found to decrease ubiquitination levels, leading to mitochondrial damage and further escalation of mitochondrial autophagy. Furthermore, in the DM-induced cell model, the upregulation of Beclin1 expression was associated with the inhibition of the ubiquitin‒proteasome system (UPS) and SKP2, thereby exacerbating autophagy. However, GBE has demonstrated notable efficacy in alleviating the reproductive toxicity induced by DM.
Our findings highlighted that SKP2 is a key regulator of Beclin1-independent autophagy and that GBE exerts therapeutic effects by upregulating SKP2 and inhibiting Beclin1 activation, which ameliorates autophagy and reduces DM-induced testicular damage.
男性不育是一个全球性的问题，与精子质量下降有关。环境污染物如溴氰菊酯（DM）对男性生殖器官有有害影响。银杏叶提取物（GBE）通过维持细胞内氧化还原稳态，可以缓解男性生殖功能障碍。然而，GBE 缓解 DM 引起的生殖毒性的机制研究有限。
本研究旨在探讨 GBE 是否通过调节 SKP2 和 Beclin1 来缓解 DM 诱导的睾丸和支持细胞生殖毒性，为开发新的治疗方法提供理论依据。
我们研究了 GBE 缓解 DM 诱导的睾丸损伤的作用，特别关注泛素化和自噬的复杂作用。
使用 ICR 雄性小鼠和 TM4 细胞系构建实验模型。通过苏木精和伊红（H&E）染色、过碘酸-Schiff（PAS）染色、超微结构观察、免疫组织化学和免疫荧光等方法观察组织、细胞和精子形态变化。测量酶和激素水平，并使用实时定量聚合酶链反应（RT-qPCR）和 Western 印迹技术检测基因和蛋白水平。
体内实验表明，DM 暴露导致性激素水平降低、生精小管直径增大和精子发生受损。同时，DM 暴露导致泛素化水平降低，导致线粒体损伤和线粒体自噬进一步加剧。此外，在 DM 诱导的细胞模型中，Beclin1 表达的上调与泛素-蛋白酶体系统（UPS）和 SKP2 的抑制有关，从而加剧了自噬。然而，GBE 已被证明在缓解 DM 引起的生殖毒性方面具有显著疗效。
我们的研究结果表明，SKP2 是 Beclin1 非依赖性自噬的关键调节剂，GBE 通过上调 SKP2 和抑制 Beclin1 激活发挥治疗作用，从而改善自噬并减轻 DM 引起的睾丸损伤。
