RELAXIN: BIOCHEMICAL AND PHYSIOLOGICAL STUDIES

在人类和其他哺乳动物的妊娠晚期， 子宫颈的稠度变软，从而使子宫颈扩张， 胎儿的快速分娩。 在难以自发的情况下， 分娩或足月引产时，坚硬的无弹性宫颈可能是 并发症的来源。 激素松弛素是一种有效的宫颈 具有临床应用潜力女性柔软剂， 家畜 关于使用松弛素作为 宫颈软化剂将在很大程度上取决于其基础研究， 对人类以外的其他物种的影响。 主要的远程 本申请的目的是更好地理解蜂窝和 分子水平的机制，其中松弛素，与 雌激素，促进怀孕大鼠子宫颈的生长和软化。 确定雌激素和松弛素对（1）组成的影响 和超微结构，以及（2）细胞内的细胞增殖 子宫颈，在妊娠第9天及之后切除大鼠卵巢 接受激素替代治疗， 孕酮、孕酮和雌激素，或孕酮、雌激素和 松弛素 将在妊娠后期（第22天）采集宫颈， 将确定雌激素和松弛素对宫颈细胞的影响 通过光学和电子显微镜。 免疫组织化学将用于 (3)确定宫颈松弛素的靶细胞。 去卵巢 妊娠大鼠也将用于研究， 测定雌激素和松弛素对酶的影响 （前列腺素合成酶、胶原酶、基质分解素） 子宫颈 子宫颈组织内的酶的量将是 通过蛋白质印迹分析和细胞定位的细胞定位， 酶将通过免疫细胞化学分析进行。 的数量 宫颈酶的mRNA将通过核糖核酸酶测定。 保护试验和酶的mRNA定位将是 通过原位杂交来完成。 本申请的具体目标5是 确定内源性松弛素是否对阴道有生理作用 怀孕的老鼠 松弛素与α-羟甲基纤维素中和的结果 单克隆抗体对大鼠松弛素的生长，拉伸性能， 并确定阴道的组织学特征。

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.