AGING EFFECTS ON TRANSVAGINAL TRANSPORT

我们广泛而长期的目标是了解调节液体和溶质在女性生殖道上皮内运动的机制。我们目前研究的主要目的是了解女性衰老如何导致宫颈粘液和阴道液的产生减少。我们将利用PI开发的一种新方法在过滤器上培养人类和宫颈/阴道上皮细胞，我们建议测试四个假设：a .衰老与宫颈和阴道上皮细胞周通透性降低有关，从而导致液体分泌减少。B.衰老使子宫颈和阴道上皮细胞中肌动蛋白丝的稳态平衡向F-肌动蛋白转移；这就产生了坚硬的细胞骨架，不允许细胞在刺激下收缩，也不能增加细胞间隙的体积，从而降低了细胞周围的通透性。与衰老相反，雌激素将平衡向g -肌动蛋白转移，从而产生更有活力的细胞骨架。C.人宫颈和阴道细胞产生一氧化氮（NO），一氧化氮能抑制肌动蛋白聚合，增加细胞周通透性。D.在人宫颈和阴道细胞中，雌激素通过上调钙依赖性NO-合成酶（ecNOS）来增加NO的产生。方法将包括在改进的扩散/使用室中进行通量和电生理研究，对附着细胞进行荧光显微镜观察以确定细胞大小的变化，生化和分子生物学分析。本研究可能为年龄和雌激素对宫颈和阴道细胞功能的影响提供新的信息。项目与健康的关系：在本研究中获得的知识可能对了解子宫颈疾病的机制很重要，例如不孕症（“宫颈因素”），以及子宫颈粘液和阴道液分泌过多或减少的情况。了解宫颈粘液产生的机制和调节对于开发改变宫颈粘液特征的化合物是重要的。这些数据可以提高我们对绝经后妇女雌激素替代的认识。它们也可以通过针对已知的经宫颈转运机制，为节育提供有效和简单的方式。

Our broad, long-term objective is to understand the mechanisms that regulate movement of fluid and solutes across the woman's reproductive tract epithelium. Our main objective in the present study is to understand how aging in women leads to decreased production of cervical mucus and vaginal fluid. We will utilize a new method that the PI has developed to culture human and cervico/vaginal epithelial cells on filters, and we propose to test four hypotheses: A. That aging is associated with decreased pericellular permeability of the cervical and vaginal epithelia to transport of fluids and solutes, which leads to decreased fluid secretion. B. That aging shifts the steady-state equilibrium of actin filaments in cervical and vaginal epithelial cells towards F- actin; this produces a rigid cytoskeleton which does not allow the cells to contract in response to stimuli and to increase the volume of the intercellular space, thus decreasing the pericellular permeability. Estrogen, in contrast to aging shifts the equilibrium towards G-actin, thus producing a more dynamic cytoskeleton. C. That human cervical and vaginal cells produce nitric oxide (NO), and that NO can inhibit actin polymerization and increase pericellular permeability. D. That in human cervical and vaginal cells, estrogen increases production of NO by up=regulation of calcium-dependent NO- synthase (ecNOS). Methods will include flux and electrophysiological studies done in a modified diffusion/ Using chamber, fluorescence microscopy of attached cells for determinations of changes in cell size, biochemical and molecular biology assays. The present study may provide novel information on age- and estrogen-related effects on cervical- and vaginal-cell function. Health relatedness of the project: the knowledge gained in present study may be important for understanding mechanisms of disease in the cervix e.g. infertility ("cervical factor"), and conditions of excessive or diminished secretion of cervical mucus and vaginal fluid. Understanding mechanisms and regulation of cervical mucus production is important for developing compounds to alter cervical mucus characteristics. These data may improve our understanding of estrogen replacement to postmenopausal women. They may also provide efficient and simple modalities for birth control by targeting known mechanisms of transcervical transport.