Mroueh J M, Arbogast L K, Fowler P, Templeton A A, Friedman C I, Danforth D R
Department of Obstetrics and Gynecology, Ohio State University, Columbus 43210, USA.
Hum Reprod. 1996 Mar;11(3):490-6. doi: 10.1093/humrep/11.3.490.
Evidence from several laboratories suggests that the ovaries of many species produce a non-steroidal factor called gonadotrophin surge-inhibiting or attenuating factor (GnSIF) which may regulate the response of the pituitary to gonadotrophin-releasing hormone (GnRH) and as such, may modulate the timing and/or amplitude of the luteinizing hormone (LH) surge. We have recently isolated a candidate GnSIF from porcine follicular fluid (PFF). Porcine GnSiF is a 69 kDa protein which has undetectable inhibin and follistatin immunological and biological activity. The present study was designed to purify and identify GnSIF from human follicular fluid. GnSIF activity was measured as suppression of GnRH-stimulated LH secretion from rat pituitary cells in primary culture. Human follicular fluid (approximately 500 ml) was recovered from patients undergoing in-vitro fertilization (IVF). GnSIF was purified by heparin-sepharose, Q-sepharose, S-sepharose, and hydrophobic interaction chromatography followed by isoelectric focusing. Gel electrophoresis and Western blot were used to identify human GnSIF and compare it with porcine GnSIF. Using these steps, we obtained a highly-purified preparation of GnSIF that manifests in-vitro bioactivity and chromatography characteristics similar to those observed for porcine GnSIF and that hybridizes with a porcine GnSIF antibody. Following treatment with human chorionic gonadotrophin/human menopausal gonadotrophin (HMG/HCG), human follicular fluid contained roughly 25% of the GnSIF (per mg protein) present in porcine follicular fluid. We conclude that GnSIF is present in human follicular fluid and may participate in the regulation of gonadotrophin secretion in this species.
来自多个实验室的证据表明，许多物种的卵巢会产生一种非甾体因子，称为促性腺激素激增抑制或减弱因子（GnSIF），它可能调节垂体对促性腺激素释放激素（GnRH）的反应，因此可能调节促黄体生成素（LH）激增的时间和/或幅度。我们最近从猪卵泡液（PFF）中分离出一种候选GnSIF。猪GnSiF是一种69 kDa的蛋白质，其抑制素和卵泡抑素的免疫和生物学活性无法检测到。本研究旨在从人卵泡液中纯化和鉴定GnSIF。GnSIF活性通过抑制原代培养的大鼠垂体细胞中GnRH刺激的LH分泌来测定。从接受体外受精（IVF）的患者中收集人卵泡液（约500 ml）。通过肝素琼脂糖凝胶、Q琼脂糖凝胶、S琼脂糖凝胶和疏水相互作用色谱，然后进行等电聚焦来纯化GnSIF。使用凝胶电泳和蛋白质印迹法鉴定人GnSIF并将其与猪GnSIF进行比较。通过这些步骤，我们获得了一种高度纯化的GnSIF制剂，其表现出与猪GnSIF相似的体外生物活性和色谱特征，并且与猪GnSIF抗体杂交。用人绒毛膜促性腺激素/人绝经期促性腺激素（HMG/HCG）处理后，人卵泡液中GnSIF的含量（每毫克蛋白质）约为猪卵泡液中的25%。我们得出结论，GnSIF存在于人卵泡液中，可能参与该物种促性腺激素分泌的调节。
