基于miRNA-221调控的DKK2/Wnt/β-catenin通路探讨补肾生精法调节少弱精子症模型大鼠生精细胞增殖、凋亡的机制研究

少弱精子症是导致男性不育的重要病因，精子质量下降与生精细胞的增殖、凋亡异常密切相关。“肾藏精，主生殖”理论与精子质量下降关系密切，补肾生精法在少弱精子症的治疗中应用广泛。前期基础及文献研究表明，补肾生精的左归丸可有效改善少弱精子症患者的精子质量，促进生精细胞增殖，抑制其凋亡，并上调少弱精子症大鼠睾丸组织中miRNA-221的表达，miRNA-221又可通过DKK2调控Wnt/β-catenin通路表达。因此，我们提出miRNA-221及其调控的DKK2/Wnt/β-catenin通路可能参与了少弱精子症的发生，左归丸可能通过该通路改善生精功能的科学假说。本研究通过构建miRNA-221沉默表达质粒，运用左归丸从体外、体内两方面验证其通过miRNA-221-DKK2/Wnt/β-catenin通路治疗少弱精子症的分子机制，为补肾生精法在男性不育症中的推广应用提供实验依据。

Oligoasthenospermia is an important cause of male infertility. The decline in sperm quality is closely related to the abnormal proliferation and apoptosis of spermatogenic cells. The theory of "Kidney accumulates sperm and masters reproduction" is closely related to the decline of sperm quality, and the method of invigorating the kidney and producing sperm is widely used in the treatment of oligoasthenospermia. Preliminary basic and literature research showed that Zuogui Pills for tonifying the kidney and producing sperm can effectively improve the sperm quality of patients with oligoasthenospermia, promote the proliferation of spermatogenic cells, inhibit their apoptosis, and up-regulate the level of miRNA-221 in the testis of oligoasthenospermia rats. Expression, miRNA-221 can regulate the expression of Wnt/β-catenin pathway through DKK2. Therefore, we propose that miRNA-221 and the DKK2/Wnt/β-catenin pathway regulated by miRNA-221 may be involved in the occurrence of oligoasthenospermia, and Zuoguiwan may improve the scientific hypothesis of spermatogenesis through this pathway. In this study, the miRNA-221 silencing expression plasmid was constructed, and Zuoguiwan was used to verify the molecular mechanism of miRNA-221-DKK2/Wnt/β-catenin pathway in the treatment of oligoasthenospermia through the miRNA-221-DKK2/Wnt/β-catenin pathway, to provide experimental basis for the popularization and application of the method of invigorating the kidney and promoting essence in male infertility.