Koutsilieris M, Niklinski W, Frenette G, Lemay A
Department of Obstetrics and Gynecology, Laval University, Quebec, Canada.
Fertil Steril. 1993 Jan;59(1):93-7.
To characterize and to purify the growth substances for mouse embryo fibroblasts (NIH/3T3) and endometrial-like cells of the peritoneal fluid of women with endometriosis.
A pool of 102 mL of peritoneal fluid (PF) collected from 24 women with laparoscopic evidence of endometriosis was dialyzed and thereafter chromatographed onto carboxymethyl (CM)-sepharose, heparin-sepharose, cartridges of C18 silica, and reverse-phase high-performance liquid chromatography (reverse-phase HPLC). Mitogenic activity throughout various steps of chromatography was monitored by the increase of the (3H)-thymidine incorporation into NIH/3T3 fibroblasts and enzymatically isolated epithelial cells of rabbit endometrium.
Peritoneal fluid of women with endometriosis contained mitogen(s) for NIH/3T3 fibroblasts that were retained on and eluted from CM-sepharose, heparin-sepharose, cartridges of C18 silica, and reverse-phase HPLC columns performed sequentially. Mitogenic material purified 2,500-fold exerted bioactivity on NIH/3T3 and the enzymatically isolated epithelial cells from rabbit endometrium.
Peritoneal fluid of women with endometriosis contains heparin-binding growth factors that can be purified by sequential chromatography on CM-sepharose, heparin-sepharose, and reverse-phase HPLC columns. The presence in the PF of potent mitogens for fibroblasts and endometrium-derived epithelial cells could play an important role in the pathogenesis of endometriosis.
鉴定并纯化用于小鼠胚胎成纤维细胞(NIH/3T3)以及子宫内膜异位症患者腹腔液中子宫内膜样细胞的生长物质。
从24例经腹腔镜检查证实患有子宫内膜异位症的女性中收集102 mL腹腔液(PF),进行透析,然后分别用羧甲基(CM)-琼脂糖凝胶、肝素-琼脂糖凝胶、C18硅胶柱以及反相高效液相色谱(反相HPLC)进行层析。在层析的各个步骤中,通过监测(3H)-胸腺嘧啶核苷掺入NIH/3T3成纤维细胞以及兔子宫内膜酶解分离的上皮细胞中的情况来监测促有丝分裂活性。
子宫内膜异位症患者的腹腔液中含有能使NIH/3T3成纤维细胞发生有丝分裂的物质,这些物质能依次保留在CM-琼脂糖凝胶、肝素-琼脂糖凝胶、C18硅胶柱以及反相HPLC柱上,并能从这些柱上洗脱下来。纯化了2500倍的促有丝分裂物质对NIH/3T3以及兔子宫内膜酶解分离的上皮细胞具有生物活性。
子宫内膜异位症患者的腹腔液中含有肝素结合生长因子,可通过依次在CM-琼脂糖凝胶、肝素-琼脂糖凝胶和反相HPLC柱上层析进行纯化。腹腔液中存在对成纤维细胞和子宫内膜来源的上皮细胞有强大促有丝分裂作用的物质,这可能在子宫内膜异位症的发病机制中起重要作用。