Leeuwenberg B R, Hudson N L, Moore L G, Hurst P R, McNatty K P
Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
J Endocrinol. 1996 Feb;148(2):281-9. doi: 10.1677/joe.0.1480281.
IGF-I was measured by RIA in plasma samples collected 8-hourly for 24 days which included two consecutive preovulatory surges of LH. In a separate study, ovarian venous blood was collected from animals undergoing ovariectomy on day 10 of the oestrous cycle, or 36 h later after being treated with prostaglandin with or without steroid-free bovine follicular fluid. Jugular venous blood samples were collected before, during and after surgery. Follicles were dissected from ovaries of these animals and sorted into categories of small, intermediate and large, non-atretic or atretic, and the follicular fluid was pooled and assayed for IGF-I. From another population of ovaries recovered from the slaughterhouse, granulosa, theca and corpora lutea were isolated, homogenized and assayed for IGF-I. Finally ovarian corpora lutea and granulosa cells were each incubated with tritiated amino acids overnight at 37 degrees C. Thereafter the tissues and media were sonicated, IGF-I extracted from the supernatant and tritiated IGF-I precipitated using a specific IGF-I antibody. The absence of any significant change in peripheral IGF-I concentrations following ovariectomy and the finding that the ovarian venous IGF-I concentrations (161 +/- 10 micrograms/l) were not significantly different from levels seen in peripheral blood (157 +/- 10 microgram/l) indicated that the ovary is not a net exporter of IGF-I. However, the ovary does synthesize IGF-I, as evidenced by granulosa and luteal synthesis, but probably not in quantities in excess of that utilized by ovarian tissues per se. Although the plasma IGF-I levels increased around the second preovulatory LH surge, the results overall indicated that the IGF-I concentrations in plasma are not strictly related to any major ovarian event during the oestrous cycle in the sheep. This view is based on the findings that the concentration of IGF-I in follicular fluid was not related to follicular health but correlated with those in peripheral plasma and that the ovarian venous concentrations did not vary between left and right ovaries irrespective of whether the ovaries contained a corpus luteum, dominant follicle or neither. Collectively, these results are consistent with the notion that IGF-I of ovarian origin fulfils an autocrine/paracrine function and does not have an endocrine role. Moreover, the results show that the concentrations of IGF-I in follicular fluid reflect those in peripheral plasma.
采用放射免疫分析法(RIA)对血浆样本中的胰岛素样生长因子-I(IGF-I)进行测定,样本每8小时采集一次,共采集24天,其中包括促黄体生成素(LH)连续两次排卵前高峰。在另一项研究中,从处于发情周期第10天接受卵巢切除术的动物身上采集卵巢静脉血,或者在使用或不使用无类固醇牛卵泡液的前列腺素处理36小时后采集。在手术前、手术期间和手术后采集颈静脉血样本。从这些动物的卵巢中解剖出卵泡,并分为小、中、大、非闭锁或闭锁等类别,将卵泡液汇集并测定其中的IGF-I。从屠宰场回收的另一批卵巢中分离出颗粒细胞、卵泡膜细胞和黄体,进行匀浆并测定其中的IGF-I。最后,将卵巢黄体和颗粒细胞分别在37℃下与氚标记的氨基酸一起孵育过夜。之后对组织和培养基进行超声处理,从上清液中提取IGF-I,并使用特异性IGF-I抗体沉淀氚标记的IGF-I。卵巢切除术后外周血IGF-I浓度无任何显著变化,且卵巢静脉IGF-I浓度(161±10微克/升)与外周血中浓度(157±10微克/升)无显著差异,这表明卵巢不是IGF-I的净输出器官。然而,卵巢确实能合成IGF-I,颗粒细胞和黄体的合成即为证明,但合成量可能不会超过卵巢组织自身的利用量太多。尽管在第二次排卵前LH高峰前后血浆IGF-I水平有所升高,但总体结果表明,绵羊发情周期中血浆IGF-I浓度与任何主要卵巢事件并无严格关联。这一观点基于以下发现:卵泡液中IGF-I浓度与卵泡健康状况无关,但与外周血浆中的浓度相关,且无论卵巢中是否有黄体、优势卵泡或两者皆无,卵巢静脉浓度在左右卵巢之间并无差异。总体而言,这些结果与卵巢来源的IGF-I发挥自分泌/旁分泌功能而非内分泌作用的观点一致。此外,结果表明卵泡液中IGF-I浓度反映了外周血浆中的浓度。