Brännström M, Woessner J F, Koos R D, Sear C H, LeMaire W J
Department of Obstetrics and Gynecology, University of Miami School of Medicine, Florida 33101.
Endocrinology. 1988 May;122(5):1715-21. doi: 10.1210/endo-122-5-1715.
We have examined the effects of a new synthetic inhibitor of mammalian tissue collagenase, CI-1 (N-[3-N-(benzyloxycarbonyl)amino-1-(R)carboxypropyl]L-leucyl-O-methyl-L- tyrosine N-methylamide; G. D. Searle SC 40827), and a general metalloproteinase inhibitor, 1,10-phenanthroline, on ovulation, as judged by the observation of follicular rupture, and on progesterone production of the perfused rat ovary. Ovaries of PMSG (20 IU)-primed rats were perfused for 21 h, and samples of medium were taken for analysis of progesterone concentration. The number of ovulations was estimated by counting the number of oocytes released into the perfusion chamber. Ovaries were stimulated with LH (0.1 micrograms/ml) plus 3-isobutyl-1-methylxanthine (IBMX; 0.2 mM), and this treatment resulted in a mean of 17.2 ovulations/treated ovary. 1,10-Phenanthroline dose-dependently inhibited ovulation, with 0, 0.2, and 12.5 ovulations/treated ovary at 1.0, 0.1, and 0.01 mM, respectively. This inhibition of ovulation closely paralleled the inhibition of extracted collagenase from uterus and ovary. However, 1,10-phenanthroline also suppressed progesterone release in a dose-dependent manner. Addition of the collagenase inhibitor (CI-1; 25 microM) 1 h after LH plus IBMX inhibited ovulation (6.3 ovulations/treated ovary). Its relatively inactive stereoisomer (CI-2; 25 microM) did not suppress ovulation (20.0 ovulations/treated ovary). CI-1 inhibited extracted uterine collagenase 50% at a concentration of 2 microM, whereas CI-2 was only 1/15th as effective. There was an 80% loss of CI-1 from the medium during the perfusions. Neither CI-1 nor CI-2 had any effect on LH plus IBMX-stimulated progesterone release. These data demonstrate that the general metalloproteinase inhibitor 1,10-phenanthroline is able to inhibit ovulation, but also inhibits steroidogenesis. The more specific inhibitor of collagenase, CI-1, can inhibit ovulation without affecting steroid production. These data indicate an important role for collagenase in the ovulatory process.
我们研究了一种新型哺乳动物组织胶原酶合成抑制剂CI-1（N-[3-N-(苄氧羰基)氨基-1-(R)羧丙基]-L-亮氨酰-O-甲基-L-酪氨酸N-甲基酰胺；G.D. Searle公司的SC 40827）和一种通用金属蛋白酶抑制剂1,10-菲咯啉对排卵（通过观察卵泡破裂来判断）以及对灌注大鼠卵巢孕酮生成的影响。用孕马血清促性腺激素（PMSG，20 IU）预处理的大鼠卵巢进行21小时的灌注，采集培养基样本分析孕酮浓度。通过计算释放到灌注室中的卵母细胞数量来估计排卵数。用促黄体生成素（LH，0.1微克/毫升）加3-异丁基-1-甲基黄嘌呤（IBMX，0.2毫摩尔）刺激卵巢，这种处理导致每个处理的卵巢平均有17.2个排卵。1,10-菲咯啉剂量依赖性地抑制排卵，在1.0毫摩尔、0.1毫摩尔和0.01毫摩尔时，每个处理的卵巢分别有0个、0.2个和12.5个排卵。这种对排卵的抑制与对子宫和卵巢中提取的胶原酶的抑制密切平行。然而，1,10-菲咯啉也以剂量依赖性方式抑制孕酮释放。在LH加IBMX刺激1小时后添加胶原酶抑制剂（CI-1，25微摩尔）可抑制排卵（每个处理的卵巢有6.3个排卵）。其相对无活性的立体异构体（CI-2，25微摩尔）不抑制排卵（每个处理的卵巢有20.0个排卵）。CI-1在2微摩尔浓度时可抑制提取的子宫胶原酶活性50%，而CI-2的效果仅为其1/15。在灌注过程中，培养基中的CI-1损失了80%。CI-1和CI-2对LH加IBMX刺激的孕酮释放均无任何影响。这些数据表明通用金属蛋白酶抑制剂1,10-菲咯啉能够抑制排卵，但也抑制类固醇生成。更特异性的胶原酶抑制剂CI-1可以抑制排卵而不影响类固醇生成。这些数据表明胶原酶在排卵过程中起重要作用。
