RXFP2介导INSL3影响小鼠睾丸引带细胞的初步机理研究

In recent years, with the increasingly severe development of the reproductive system and the deepening of people's understanding of it, the development of testis as the core organ has attracted much attention.The development of testis is accompanied by two descending processes, and its abnormal development is closely related to the decline.Previous studies have shown that insulin-like factor 3 (INSL3) plays a leading role in the first transabdominal phase of testicular descent, and is closely related to the important guiding structure: the development of gubernaculum. Our previous studies have confirmed INSL3 specific receptors: relaxin family peptide receptor 2 (RXFP2) exists in mice gubernaculum testis cells (GTCs), INSL3 can affect the morphology and proliferation and contraction of the GTCs, and is closely related to RXFP2. At present, the understanding of RXFP2 is still limited, and the mechanism of INSL3 affecting the development of testis is also unclear. Peptide hormone binded its receptors could produce biological effect through signal transduction pathway. So, we explore the value of RXFP2 in the mechanism of INSL3 influence mice gubernaculum cell development via PKA and PKC signaling pathway in vitro.

近年来，随着生殖系统发育异常形势的日益严峻和人们对之认识的逐步加深，睾丸作为核心器官其发育备受关注。睾丸的发育伴随着两个下降过程，其异常发育与下降密切关联。既往研究表明胰岛素样因子3（INSL3）在睾丸经腹下降期起主导作用，且与其中重要的引导性结构：睾丸引带的发育关系密切。我们前期研究也证实INSL3特异性受体：松弛素家族肽受体2（RXFP2）存在于小鼠睾丸引带细胞（GTCs）上，INSL3可影响GTCs的形态结构及增殖和收缩活性，且与RXFP2密切相关。目前对RXFP2的了解仍有限，对INSL3影响睾丸引带发育的作用机理也很不清楚。结合肽类激素可直接作用于其受体，激活胞内第二信号系统而发挥生物学效应的机理研究，本项目拟通过培养GTCs，基于PKA和PKC信号途径探讨RXFP2介导INSL3影响GTCs发育的可能机制，以深化INSL3及RXFP2在睾丸下降甚至整个雄性生殖系统发育中的作用。

睾丸下降不全是小儿最常见的先天性泌尿生殖系统疾病。睾丸下降发育不全可影响日后性和生殖能力，甚至增加罹患肿瘤机率，使得深入研究睾丸下降发育的影响因素及机制尤为重要。.本项目从颇富特色的睾丸引带角度研究睾丸下降发育的机理。通过本研究发现，RXFP2可能是引起小鼠睾丸引带细胞异常分化的潜在分子标志物。在具有组织细胞特异性和Crosstalk（交互应答）等复杂机制的细胞信号通路网的研究中，我们首先探究PKA和PKC信号通路在INSL3影响小鼠睾丸引带细胞增殖及分化中所起的作用。本项目通过分子生物学等技术研究发现，INSL3对小鼠睾丸引带细胞有促进增殖及抑制凋亡作用，PKA和PKC细胞信号通路参与了INSL3对引带细胞的促增殖及抗凋亡作用，INSL3可上调小鼠睾丸引带细胞PKA和PKC蛋白磷酸化，提示INSL3通过激活PKA/PKC信号通路调节小鼠睾丸引带细胞的的增殖和凋亡。从信号通路的分子水平进一步揭示了睾丸引带发育的机制。.通过本研究，我们确定了RXFP2是参与调控睾丸下降过程中睾丸引带发育的关键基因，在INSL3介导睾丸引带发育的信号转导中起重要作用；并进一步证明了PKA/PKC信号通路参与了INSL3/RXFP2调控小鼠睾丸引带细胞的增殖、迁移和凋亡等生物学活动。我们的发现揭示了INSL3/RXFP2调节小鼠睾丸引带发育的潜在机制，为深入揭示睾丸下降发育的机理增加新的信息。有望为先天性睾丸下降发育不全的产前筛查提供一个新的潜在生物标志物，也可能为睾丸下降不全的预防和临床非手术治疗提供新的思路和治疗靶点。.