Hynes A C, Kane M T, Sreenan J M
Department of Physiology, University College, Galway, Ireland.
J Reprod Fertil. 1996 Nov;108(2):185-91. doi: 10.1530/jrf.0.1080185.
Bovine follicular fluid was aspirated from follicles of 2-20 mm in diameter, charcoal-treated to remove steroids and then separated into low and high molecular mass fractions. The low molecular mass (< 10 kDa) fraction was purified on a Sephadex G-25 chromatography column with formic acid as the eluent. Seven peaks were isolated and assayed for biological activity in cultures of bovine granulosa cells at concentrations of 10, 100 and 1000 ng ml-1. One peak (peak 4) inhibited (P < 0.001) the proliferation of granulosa cells when measured by cell counting and by [3H]thymidine incorporation (33-37% inhibition). This peak inhibited proliferation of granulosa cells from both small (< 2 mm) and medium (2-10 mm) follicles, but not large (> 10 mm) follicles. The inhibitory effect of peak 4 was not due to a toxic effect on cells. Administration of peak 4 to rats did not affect liver or kidney masses but did decrease uterine (25%, P < 0.01) and ovarian (35%, P < 0.01) masses. Peak 4 also caused a reduction in the number of large follicles (65%, P < 0.01) but increased the number of small follicles (55%, P < 0.01). We have named the inhibitory factor associated with peak 4, granulosa cell-inhibitory factor (GCIF). The results presented suggest that GCIF may be a factor secreted by dominant follicles that inhibits the development of subordinate follicles.
从直径为2 - 20毫米的卵泡中抽吸牛卵泡液，经活性炭处理以去除类固醇，然后分离为低分子量和高分子量组分。低分子量（<10 kDa）组分在以甲酸为洗脱剂的Sephadex G - 25色谱柱上进行纯化。分离出七个峰，并在牛颗粒细胞培养物中以10、100和1000 ng ml-1的浓度测定其生物活性。通过细胞计数和[3H]胸苷掺入法测量时，一个峰（峰4）抑制（P < 0.001）颗粒细胞的增殖（抑制率为33 - 37%）。该峰抑制来自小（<2毫米）和中（2 - 10毫米）卵泡的颗粒细胞增殖，但不抑制大（>10毫米）卵泡的颗粒细胞增殖。峰4的抑制作用并非由于对细胞的毒性作用。给大鼠注射峰4不影响肝脏或肾脏重量，但确实使子宫重量减少（25%，P < 0.01）和卵巢重量减少（35%，P < 0.01）。峰4还导致大卵泡数量减少（65%，P < 0.01），但小卵泡数量增加（55%，P < 0.01）。我们将与峰4相关的抑制因子命名为颗粒细胞抑制因子（GCIF）。所呈现的结果表明，GCIF可能是优势卵泡分泌的一种抑制从属卵泡发育的因子。
