瘦素对附睾精子成熟的影响及机制研究

Male infertility has a great adverse effect on human reproductive health. Whereas, for the most male infertility patients with sperm maturation dysfunction, it is difficult to identify its causes and the specific mechanisms. In order to safeguard and promote male reproductive health, it is of great importance to further study the mechanism of epididymal sperm maturation. Our previous studies found that abnormal expression of seminal Leptin was closely related to asthenospermia, implying that Leptin plays an important role in the regulation of epididymal sperm maturation. However, the molecular mechanism of the regulation is still not clear, the pilot experiments showed some crosstalk with the signaling pathway of TNF-α and IL-6. Therefore, in order to identify the impact of Leptin on sperm maturation in this project, we will do the research on three levels, anmianal experiments ,cell experiments and human experiments. On each level, we will observe the changes of Leptin and its receptor expressions on caput and cauda epididymides, epitaphial cell function and proliferation, expressions of inflammatory factors, sperm motility, membrane function and ultrastructure. Firstly, on the animal level, we will construct different animal models, whose expression of Leptin signaling pathway or inflammatory factors are abnormal, intend to clarify the impact of Leptin on epididymal sperm maturation and the role of inflammatory factors. Secondly, on the cell level, we will directly intervene epididymal epithelia and epdidymal sperm by Leptin and make antagonistic biological effects through different signaling pathway inhibitors, in order to explore the specific regulation and signaling pathways of Leptin. Finally, on the human level, we will analysis patients with asthenospermia or genital tract infection, intend to verify the relation of Leptin and human epididymal sperm maturation. Thus, our project will reveal the repercussion of Leptin on sperm maturation for the sake of providing a potential new target for the treatment to the male infertility patients with sperm maturation dysfunction.

相当一部分男性不育症患者存在精子成熟障碍，深入研究影响精子成熟障碍的原因和机制对于诊断与治疗男性不育症具有重要意义。我们前期工作发现精浆瘦素表达异常与弱精子症等精子成熟障碍密切相关，但其具体机制尚未明确，初步研究结果提示可能与IL-6及TNF-α的信号通路有关。本项目包括动物水平、细胞水平和人体水平三个层次的研究，分别观察瘦素及其受体在附睾头部和尾部表达的变化，炎症因子表达的变化，附睾上皮细胞功能、增殖分化凋亡的变化，精子活力、膜功能及超微结构的变化。首先，在动物水平，通过构建瘦素信号通路异常动物模型和附睾炎模型进行研究，明确瘦素对附睾精子成熟的影响以及炎症因子在其中的作用。其次，在细胞水平，直接进行瘦素及信号通路拮抗剂干预的研究，阐明其具体调控机制及信号通路。最后在人体水平，对生殖道感染、弱精子症等男性不育症患者进行分析，验证瘦素对人类附睾精子成熟的作用，为治疗男性不育症提供潜在新靶点。

深入研究影响精子成熟障碍的原因和机制对于诊断与治疗男性不育症具有重要意义。我们前期工作发现精浆瘦素表达异常与弱精子症等精子成熟障碍密切相关，但其具体机制尚未明确。本项目包括动物水平、细胞水平和分子机制和人体水平四个层次的研究：首先动物水平上，通过构建动物模型明确 对附睾的影响，具体而言，通过高脂饮食构建瘦素高表达大鼠模型，探索瘦素对附睾分泌功能影响的在体（in vivo）机制。随后，通过构建瘦素高表达大鼠模型研究瘦素影响附睾功能的在体机制。第三，构建大鼠附睾炎动物模型，检测附睾组织炎症因子IL-1β，TNF-α，IL-6表达，检测附睾组织及受体Ob-R表达级定位改变。大鼠精子活力，活率，形态检测。最后，转录组测序分析了附睾炎时信号调控网络的改变，为附睾炎的机制研究和相关基因研究提供了方向，也为后续与附睾炎之间的研究提供了参考意义。在细胞水平上，我们研究 对附睾精子成熟的影响及调控机制，包括 对附睾上皮细胞的影响及调控机制， 对附睾中精子的影响及调控机制。.随后，我们聚焦于 与炎症因子 TNF-α、IL-6 的相互作用对附睾的影响及分子机制。构建附睾炎动物模型，比较观察各组 及其受体在附睾组织表达情况，相关炎症因子表达情况，附睾组织功能、增殖、分化及凋亡变化情况。附睾上皮细胞分离培养，研究与炎症因子 TNF-α、 IL-6 的相互作用对附睾的影响及分子机制, 对附睾上皮细胞的影响及调控机制，瘦素与附睾上皮细胞炎症及对 NF-κB调控炎症作用的研究。人体水平上研究 对附睾精子成熟的影响，在弱精子症男性不育患者等精子成熟障碍患者，生殖道感染男性不育患者、白细胞精液症患者、男性不育症伴代谢综合症患者等炎症因子高表达患者，验证瘦素对人类附睾精子成熟的作用，为治疗男性不育症提供潜在新靶点。