The effect of nonoxynol-9 on human endometrium

描述（由申请人提供）：避孕杀菌剂提供了妇女控制避孕和预防艾滋病毒的可能性。一份报告称，多次服用壬氧醇醚-9 （N-9）后，艾滋病毒感染率增加，这引起了严重的公共卫生关切。这一发现可能与阴道上皮破坏和HIV宿主炎症细胞募集有关，因为生殖器毒性。几条线索的证据支持存在生理性液体运动从阴道到上生殖道通过子宫蠕动。因此，必须确定候选杀微生物剂对子宫内膜和上生殖道的毒性。在南加州大学进行的一项初步研究中，用三剂量的N-9处理子宫内膜外植体，显示出对子宫内膜完整性的多重毒性作用。我们通过免疫组织化学证明，M30抗体染色显示早期凋亡的剂量相关增加，DNA片段化显示所有剂量的N-9都存在晚期凋亡。此外，我们证明了子宫内膜炎症反应的消除和muc1表达的剂量依赖性降低，muc1是一种在子宫内膜上皮上形成保护性粘蛋白层的蛋白质。本研究的目的是确定N-9在体内和体外对人子宫内膜在月经周期的两个阶段的影响。28名妇女将接受一个治疗前周期，在此期间将进行子宫冲洗和子宫内膜活检。受试者将使用N-9涂抹1 - 3天，并进行第二次子宫灌洗和活检。将检查灌洗液是否有细胞因子和炎症细胞。子宫内膜活检组织将使用光镜和电子显微镜进行评估，并进行M30 CytoDeath抗体和MUC-1染色。提取DNA进行凝胶电泳。预处理活检组织将作为外植体培养，并进行平行评估。本研究结果将为杀微生物剂对子宫内膜毒性的作用提供重要的初步信息，并可能有助于指导未来的研究重点。

DESCRIPTION (provided by applicant): Contraceptive microbicides offer the possibility of women-controlled contraception and HIV prevention. Serious public health concerns were raised following a report that HIV infection rates increased after multiple doses of nonoxynol-9 (N-9). This finding may be related to vaginal epithelium disruption and recruitment of HIV host inflammatory cells because of genital toxicity. Several lines of evidence support the existence of physiologic fluid movement from the vagina to the upper reproductive tract by uterine peristalsis. Therefore, the toxicity of microbicidal candidates on the endometrium and upper reproductive tract must be determined. A preliminary study performed at USC using an endometrial explant treated with three doses of N-9, showed multiple toxic effects on endometrial integrity. We demonstrated, using immunohistochemistry, a dose-related increase in early apoptosis by M30 antibody staining, and presence of late apoptosis at all doses of N-9 by DNA fragmentation. In addition, we demonstrated obliteration of the endometrial inflammatory response and a dose-dependent reduction in expression of MUC-1, a protein that creates a protective mucin layer over endometrial epithelium. The purpose of the current study is to determine the effects of N-9 on human endometrium in vivo and in vitro during both phases of the menstrual cycle. Twenty-eight women will undergo a pre-treatment cycle during which uterine lavage and endometrial biopsies will be performed. Subjects will then apply N-9 for one or three days and have a second uterine lavage and biopsy. Lavage fluids will be examined for cytokines and inflammatory cells. Endometrial biopsy tissue will be evaluated using light and electron microscopy, and stained for M30 CytoDeath antibody and MUC-1. DNA will be extracted for gel electrophoresis. Tissue from pre-treatment biopsies will be cultured as explants and parallel evaluations performed. The results of this study will provide important preliminary information on the role of microbicides on endometrial toxicity and may help guide future research priorities.