强精煎对实验性大鼠精子发生的影响及其调控机制的研究

Oligoasthenozoospermia is the main clinical manifestation of male infertility, as the research hotspot in recent years, the concrete mechanism of spermatogenesis and sperm maturation still has not been classified, our previous study indicated that the Qiang Jing Docoction has a great clinical efficacy, it can significantly improve sperm motility and sperm count, repair pathological damage of the testicles of the model rats with oligoasthenozoospermia, it also has anti-spermatogenic cell apoptosis effcts and antioxidant effects.This subject is based on the early theoretical and experimental research, starting from the factors like the Qiang Jing Docoction might activates the proliferation and differentiation of the spermatogonial cell,maintains the homeostasis in the process of spermatogenesis, improves the capacity of spermatogenic cell and sperm mitochondria by regulating c-kit and CFTR.We tend to learn about the effects and regulation mechanism on spermatogenesis of experimental rat of Qiang Jing Docoction, to provide experimental basis for promoting the application of Qiang Jing Docoction treating male infertility, and to provide theoretical basis for traditional Chinese medicine treating male infertility. Also, through this study, we tend to provide new basis and ideas for understanding the pathological mechanism of the spermatogenesis regulation and male infertility,and to enrich the"Spleen - mitochondrial related theory"further.

少弱精子症是男性不育症的主要临床表现之一，精子发生与成熟的调控机制尚未完全阐明，是近年研究热点之一，我们的前期研究表明，中药强精煎临床能显著改善精子数量和活力，能显著修复少弱精子症模型大鼠的睾丸病理损伤、具有抗生精细胞凋亡及抗氧化作用。该课题基于前期理论和实验研究基础，从强精煎可能通过调控睾丸c-kit和CFTR，启动精原细胞的增殖分化，维持精子发生发育过程中微环境的稳定，增强生精细胞及精子线粒体功能，提升精子受精能力等因素入手，重点探讨强精煎调控实验性大鼠精子发生发育的作用机制，为推广应用强精煎治疗男性不育症提供新的实验依据，尤其是为中医药在男性不育症中应用提供理论依据,为理解精子发生的调控和男性不育症的病理机理提供新的依据和思路,进一步充实丰富"中医脾-线粒体相关学说"。

高达60％～75％的男性不育症原因不明，表现为少精、弱精和/或畸形精子症等精子质量异常。生精功能障碍是导致男性不育主要原因之一。c-kit在调控精原干细胞分化的机制中具有决定性作用。囊性纤维化跨膜转导调节因子（CFTR）突变频率的增加或表达减少与男性先天性输精管缺如 、精子畸形，无精子症和性腺等双边缺失相关。CFTR蛋白在精子中有大量表达，在精子获能和受精中起关键作用。线粒体能量代谢是细胞功能的一个关键因素，也是生殖细胞发育和受精过程关键代谢途径。线粒体的损伤可引起细胞凋亡，最终导致精子生成减少。本课题从调控生精和受精能力的分子生物学机制入手进行研究，以黄精赞育胶囊作阳性对照，观察强精煎对生精障碍模型大鼠睾丸/精子c-kit、CFTR蛋白和基因、线粒体Na+－K+－ATP酶、Mg2+-ATP酶及Ca2+－ATP酶活性、线粒体膜电位、生精细胞和精子超微结构的干预效应。结果：与模型组相比，强精煎低剂量、高剂量干预后的模型大鼠精子数量及活力均优于模型组，睾丸组织/精子Na+－K+－ATP酶、Mg2+-ATP酶及Ca2+－ATP酶活性均优于模型组，线粒体膜电位正常率和荧光强度均高于模型组；模型组睾丸、精子c-kit和CFTR蛋白/基因表达明显下调，药物干预后c-kit和CFTR蛋白/基因表达明显上调。常规病理提示模型组生精上皮明显破坏，药物干预后得到不同程度恢复，超微病理进一步观察见模型大鼠生精细胞线粒体大量减少，结构模糊或缺失，其余细胞器也减少。强精煎两组和黄精赞育胶囊组大鼠生精细胞线粒体等细胞器较为丰富，无明显破坏（强精煎低剂量组线粒体较多呈肿胀状态），精子尾部微管9+2结构紧密。结果表明，强精煎可通过增强线粒体酶活性，改善线粒体膜电位，改善能量代谢，修复生精上皮及其亚细胞结构，保护生精功能，改善精子活力；上调c-kit和CFTR蛋白/基因表达等多途径、多靶点作用调控精子发生而达到提高精子数量，改善精子活力以治疗少、弱精子症的目的，为临床用于治疗少、弱精子症/生精功能障碍提供了实验依据，为中医药治疗男性不育症提供理论依据。

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