AGING EFFECTS ON TRANSVAGINAL TRANSPORT

我们广泛的、长期的目标是了解调节女性生殖道上皮中液体和溶质运动的机制。我们在这项研究中的主要目标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和宫颈/阴道上皮细胞，并建议检验四个假设：A.衰老与宫颈和阴道上皮细胞周对液体和溶质运输的渗透性降低有关，这导致液体分泌减少。B.衰老使宫颈和阴道上皮细胞肌动蛋白细丝的稳态平衡转向F-肌动蛋白；这产生了一种坚硬的细胞骨架，使细胞不能对刺激做出反应而收缩，并增加了细胞间隙的体积，从而降低了细胞周通透性。与衰老不同的是，雌激素将平衡转向G-肌动蛋白，从而产生更具活力的细胞骨架。C.人类宫颈和阴道细胞产生一氧化氮(NO)，NO可以抑制肌动蛋白聚合并增加细胞周通透性。在人类宫颈和阴道细胞中，雌激素通过上调钙依赖的一氧化氮合酶(EcNOS)来增加一氧化氮的产生。方法将包括在改进的扩散/使用小室中进行的助熔剂和电生理研究，用于确定细胞大小变化的附着细胞的荧光显微镜，生物化学和分子生物学分析。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。项目的健康相关性：本研究中获得的知识对于了解宫颈疾病的机制可能很重要，例如不孕症(“宫颈因素”)，以及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性的化合物很重要。这些数据可能会提高我们对绝经后妇女雌激素补充的理解。它们还可以通过针对已知的宫内转运机制提供有效和简单的节育方法。

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.