ENDOMETRIOSIS-ASSOCIATED SECRETORY PROTEINS

子宫内膜异位症是一种引起衰弱疼痛和 女性的生育能力。尽管有这些毁灭性的影响，但这种机制通过 目前尚不清楚这种神秘的疾病是如何发挥这些致病作用的。 虽然组织学上与子宫内膜相似，但子宫内膜异位症 组织在生物化学上与子宫对应组织不同 各式各样的时装。细胞的生化特性发生改变 子宫内膜异位症组织可能与免疫系统的异常有关， 排卵、受精和植入都有牵连 作为子宫内膜异位症相关性不孕症的原因。 使用大鼠子宫内膜异位症模型，两个以前未知的 子宫内膜异位症相关蛋白已被鉴定。黄体酮- 诱导的子宫蛋白PUP-1可能在正常生殖中发挥作用 包括植入在内的过程。奇怪的是，合成和/或分泌 子宫内膜异位植入物中PUP-1的表达与之24小时不同 子宫内膜的组织结构。第二种蛋白质EP-1是通过 子宫内膜异位症，但不是子宫内膜组织。EP-1可能是一个有用的标志物 在子宫内膜异位症的发病机制中起一定作用。 PUP-1和EP-1最近在培养基中被鉴定为 纯化的大鼠和人子宫内膜及异位内膜种植基质 细胞。 该提案目标是提纯和表征老鼠和 为了洞察人类PUP-1和EP-1的结构、模式 这些蛋白质的表达和生理功能。具体的 本项目的目标是：1)开发PUP-1的纯化方案 和EP-1；2)产生a)抗血清，b)放射免疫测定和c)限制 PUP-1和EP-1的蛋白质序列数据；3)分子克隆 代表PUP-1和EP-1，并获得核苷酸序列数据。 蛋白质纯化将采用细胞培养和层析技术 技巧。抗血清、放射免疫分析和蛋白质序列数据 从纯化的蛋白质中提取的蛋白质将用于检测和监测 PUP-1和EP-1在不同组织和血清中的分布 妊娠早期和服用类固醇后的生殖周期 并获得PUP-1和EP-1的DNA序列数据。这个 核酸序列数据将与其他序列信息进行比较 在数据库中提供对身份的进一步洞察和可能 蛋白质的功能。 这些研究将支持长期目标，包括确定 PUP-1和EP-1在生殖功能和病理生理中的作用 子宫内膜异位症。PUP-1和EP-1的特性也可能导致 无创性血清黄体酮标志物的发展- 子宫内膜异位症与子宫内膜功能的相关性。 最终，这些研究可能会改善诊断、预后和 子宫内膜异位症的治疗方法。

Endometriosis is a disease that causes debilitating pain and loss of fertility in women. Despite these devastating effects, the mechanism by which this enigmatic disease exerts these pathogenicities is unknown. Although histologically similar to uterine endometrium, endometriotic tissue is biochemically different from its uterine counterpart in a variety of fashions. The altered biochemical characteristics of the endometriotic tissue may be involved with anomalies of the immune system, ovulation, fertilization, and implantation which have all been implicated as causes of endometriosis-associated infertility. Using a rat model for endometriosis, two previously unknown endometriosis-associated proteins have been identified. A progesterone- induced uterine protein, PUP-1, may play a role in normal reproductive processes including implantation. Curiously, synthesis and/or secretion of PUP-1 by the endometriotic implant is 24 hours out of phase with that of the uterine endometrium. A second protein, EP-1, is synthesized by endometriotic but not endometrial tissue. EP-1 may be a useful marker of endometriosis and have a role in the pathophysiology of the disease. PUP-1 and EP-1 have recently been identified in culture media from purified rat and human endometrial and endometriotic implant stromal cells, respectively. The goal of this proposal is to purify and characterize both rat and human PUP-1 and EP-1 in order to obtain insight into the structure, mode of expression and physiological function of these proteins. The specific aims of this project are to: 1) develop a purification scheme for PUP-1 and EP-1; 2) generate a) antisera, b) radioimmunoassays and c) limited protein sequence data for PUP-1 and EP-1; and 3) molecularly clone cDNA representing PUP-1 and EP-1 and obtain nucleotide sequence data. Protein purification will employ cell culture and chromatographic techniques. the antisera, radioimmunoassays and protein sequence data derived from the purified proteins will be used to detect and monitor the distribution of PUP-1 and EP-1 in various tissues and in sera throughout the reproductive cycle, during early pregnancy and following steroid treatment as well as to obtain DNA sequence data for PUP-1 and EP-1. The nucleic acid sequence data will be compared to other sequence information in data banks to provide further insight into the identity and possible function of the proteins. These studies will support long term goals which include determining the role of PUP-1 and EP-1 in reproductive function and the pathophysiology of endometriosis. The characterization of PUP-1 and EP-1 may also lead to the development of non-invasive serum markers for progesterone- dependent endometrial function and the disease endometriosis. Ultimately, these studies may lead to improved diagnostic, prognostic and therapeutic methods for the management of endometriosis.