RELAXIN: BIOCHEMICAL AND PHYSIOLOGICAL STUDIES

在人类和其他哺乳动物的妊娠晚期， 宫颈的一致性变软，从而使宫颈扩张和 胎儿的快速分娩。在难以自发性的情况下 足月分娩或引产，硬而无弹性的宫颈可能是 并发症的根源。松弛激素是一种有效的宫颈 具有临床应用潜力的女性和女性柔软剂 家养动物。关于松弛素AS使用的知情决定 宫颈软化剂将在很大程度上取决于对其的基础研究 对人类以外的物种的影响。主力远程 此应用程序的目标是更好地了解蜂窝和 分子水平松弛的机制(S)，松弛与 雌激素，促进怀孕大鼠宫颈的生长和软化。 测定雌激素和松弛素对(1)成分的影响 和超微结构，以及(2)细胞内细胞的增殖 子宫颈，大鼠将在怀孕第9天及以后切除卵巢 接受激素替代治疗，生理水平为 黄体酮、黄体酮和雌激素，或黄体酮、雌激素和 放松药。子宫颈将在怀孕后期(第22天)收集， 雌激素和松弛素对宫颈细胞的影响将被确定 通过光学和电子显微镜观察。免疫组织化学将用于 (3)确定宫颈松弛素的靶细胞。卵巢摘除 荷尔蒙治疗的怀孕大鼠也将用于旨在 雌激素和松弛素对酶活性影响的测定 大鼠合成的前列腺素合成酶、胶原酶、基质分解素 子宫颈。宫颈组织中的酶的数量将是 通过Western印迹分析和细胞定位确定 酶将通过免疫细胞化学分析来完成。数量上的 宫颈酶的mRNA将通过核糖核酸酶来测定。 酶的保护实验和信使核糖核酸的定位 通过原位杂交完成。本申请的具体目标5是 确定内源性松弛素对阴道是否有生理作用 在怀孕的老鼠身上。松弛素与阿司匹林中和的后果 抗大鼠松弛素单抗对生长、拉伸性能的影响 阴道的组织学特征将被确定。

During late pregnancy in humans and other mammalian species, the consistency of the cervix softens, thus enabling cervical dilatation and rapid delivery of the fetus(es). In cases of difficult spontaneous delivery or induced delivery at term, a hard inelastic cervix may be a source of complications. The hormone relaxin is an effective cervical softening agent with potential for clinical application in women and domestic animals. Informed decisions concerning the use of relaxin as a cervical softening agent will largely depend upon basic studies of its effects in species other than the human. The principal long-range objective of this application is to better understand at the cellular and molecular levels the mechanism(s) whereby relaxin, in concert with estrogen, promotes growth and softening of the cervix in pregnant rats. To determine the influence of estrogen and relaxin on (1) the composition and ultrastructure, as well as (2) the proliferation of cells within the cervix, rats will be ovariectomized on day 9 of pregnancy and thereafter receive hormone replacement therapy with physiological levels of progesterone, progesterone and estrogen, or progesterone, estrogen, and relaxin. Cervices will be collected late in pregnancy (day 22) and the effects of estrogen and relaxin on the cervical cells will be determined by light and electron microscopy. Immunohistochemistry will be used to (3) identify the target cells for relaxin in the cervix. Ovariectomized hormone-treated pregnant rats will also be used for studies aimed at determining the influence of estrogen and relaxin on enzymes (prostaglandin synthase, collagenase, stromelysin) synthesized by rat cervices. The quantities of enzymes within cervical tissues will be determined by Western blot analysis and the cellular localization of the enzymes will be done by immunocytochemical analysis. The quantities of mRNA for the cervical enzymes will be determined by ribonuclease protection assay and the localization of the mRNA for enzymes will be done by in situ hybridization. Specific Aim 5 of this application is to determine if endogenous relaxin has physiological effects on the vagina in pregnant rats. The consequences of neutralization of relaxin with a monoclonal antibody for rat relaxin on the growth, tensile properties, and histological characteristics of the vagina will be determined.