Kennedy J L, Muse K N, Keeble S C, Curry T E
Devision of Reproductive Endocrinology, Department of Obstetrics and Gynecology, University of Kentucky Medical Center, 800 Rose Street, 40536-0084, Lexington, KY.
Endocrine. 1996 Dec;5(3):299-305. doi: 10.1007/BF02739063.
Tissue inhibitors of metalloproteinases (TIMPs) are membres of a multigene family of proteinase inhibitors that regulate the activity of metalloproteinases. To test the hypothesis that TIMPs regulate connective tissue remodeling during follicular development, rats were injected with PMSG (20 IU, sc), and ovaries and serum were collected at the time of pregnant mare serum gonadotropin at the time of pregnant mare serum gonadotropin (PMSG) administration (0 h) and at 6, 12, 24, 36, and 48 h later for analysis of TIMP expression, metalloproteinase inhibitor activity, and steroidogenesis. Serum estradiol levels increased from 20.9 pg/mL at 0 h to 461 pg/mL at 48 h. Northern analysis was performed for analysis of TIMP-1, TIMP-2, and TIMP-3 expression (N = 4). For TIMP-1, PMSG stimulated a 2.4- to 2.5-fold increase in TIMP-1 mRNA at 6 and 12 h compared to ovaries collected at the time of PMSG administration (i.e., 0 h control). TIMP-1 mRNA returned to control levels within 24 h and remained unchanged through 48 h. In contrast to TIMP-1, TIMP-3 mRNA decreased by approx 2.5-fold at 6 h following PMSG administration, and expression remained decreased through 48 h. For TIMP-2, the expression of the 3.5-kb transcript decreased at 24 h after PMSG, whereas expression of the 1 kb transcript was unchanged. There was no change in metalloproteinase inhibitor activity in whole ovarian extracts between 0 and 36 h. However, there was an increase in inhibitor activity at 48 h. These findings are the first demonstration of hormonal regulation of TIMPs during the follicular phase. The differential regulation of the TIMPs by gonadotropins, for example, an increase in TIMP-1 and a concomitant decrease in TIMP-3 expression, may reflect different roles, sites of action, or enzyme specificity for the inhibitors as the follicle grows.
金属蛋白酶组织抑制剂（TIMPs）是调节金属蛋白酶活性的蛋白酶抑制剂多基因家族的成员。为了验证TIMPs在卵泡发育过程中调节结缔组织重塑的假说，给大鼠注射孕马血清促性腺激素（PMSG，20 IU，皮下注射），并在注射孕马血清促性腺激素（PMSG）时（0小时）以及之后6、12、24、36和48小时采集卵巢和血清，用于分析TIMP表达、金属蛋白酶抑制剂活性和类固醇生成。血清雌二醇水平从0小时的20.9 pg/mL增加到48小时的461 pg/mL。进行Northern分析以分析TIMP-1、TIMP-2和TIMP-3的表达（N = 4）。对于TIMP-1，与PMSG给药时（即0小时对照）采集的卵巢相比，PMSG在6和12小时刺激TIMP-1 mRNA增加2.4至2.5倍。TIMP-1 mRNA在24小时内恢复到对照水平，并在48小时内保持不变。与TIMP-1相反，PMSG给药后6小时TIMP-3 mRNA下降约2.5倍，并且表达在48小时内持续下降。对于TIMP-2，3.5 kb转录本的表达在PMSG后24小时下降，而1 kb转录本的表达未改变。在0至36小时之间，整个卵巢提取物中的金属蛋白酶抑制剂活性没有变化。然而，在48小时时抑制剂活性增加。这些发现首次证明了卵泡期TIMPs的激素调节。促性腺激素对TIMPs的差异调节，例如TIMP-1增加而TIMP-3表达同时下降，可能反映了随着卵泡生长，抑制剂的不同作用、作用位点或酶特异性。
