Abstract：
Testosterone, an important sex hormone, regulates sexual maturation, testicular development, spermatogenesis and the maintenance of secondary sexual characteristics in males. Testicular Leydig cells are the primary source of testosterone production in the body. Hezuo pigs, native to the southern part of Gansu, China, are characterized by early sexual maturity, strong disease resistance and roughage tolerance. This study employed type IV collagenase digestion combined with cell sieve filtration to isolate and purify Leydig cells from the testicular tissue of 1-month-old Hezuo pigs. We also preliminarily investigated the functions of these cells. The results indicated that the purity of the isolated and purified Leydig cells was as high as 95%. Immunofluorescence analysis demonstrated that the isolated cells specifically expressed the 3β-hydroxysteroid dehydrogenase antibody. Enzyme-linked immunosorbent assay results showed that the testosterone secretion of the Leydig cells cultured in vitro (generations 5-9) ranged between 1.29-1.67 ng/mL. Additionally, the content of the cellular autophagy signature protein microtubule-associated protein 1 light chain 3 was measured at 230-280 pg/mL. Through this study, we established an in vitro system for the isolation, purification and characterization of testicular Leydig cells from 1-month-old Hezuo pigs, providing a reference for exploring the molecular mechanism behind precocious puberty in Hezuo pigs.
摘要：
睾酮,一种重要的性激素,调节性成熟，睾丸发育,男性的精子发生和第二性征的维持。睾丸间质细胞是体内睾酮产生的主要来源。合作猪,原产于甘肃南部，中国,以性成熟早为特征，抗病性强，耐粗饲料。本研究采用IV型胶原酶消化结合细胞筛过滤，从1月龄合作猪的睾丸组织中分离纯化睾丸间质细胞。我们还初步研究了这些细胞的功能。结果表明，分离纯化的Leydig细胞的纯度高达95％。免疫荧光分析表明，分离的细胞特异性表达了3β-羟基类固醇脱氢酶抗体。酶联免疫吸附试验结果显示,体外培养的Leydig细胞(第5-9代)的睾酮分泌在1.29-1.67ng/mL之间。此外,细胞自噬特征蛋白微管相关蛋白1轻链3的含量为230-280pg/mL.通过这项研究,我们建立了一个体外分离系统，1月龄合作猪睾丸间质细胞的纯化和鉴定,为探索合作猪性早熟的分子机制提供参考。
