H AND HC03 TRANSPORT BY AIRWAY EPITHELIA IN CF

H 和 HC03 CF 中的气道上皮运输

• 批准号: 6680920
• 负责人: TERRY E MACHEN
• 金额: $ 22.57万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-09-30 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6680920
• 关键词: Golgi apparatus; acid base balance; bicarbonates; chloride channels; cystic fibrosis; electrophysiology; fluorescence microscopy; human tissue; hydrogen transport; ion transport; membrane permeability; mucus; respiratory epithelium; secretion; sodium channel; tight junctions; tissue /cell culture; western blottings

DESCRIPTION (Applicant's Abstract): Acid - base transport by airway epithelial cells likely play key roles in cystic fibrosis (CF) due to effects on mucus hydration and rheology, ciliary transport, bacterial binding and anti-microbial factors. Airway surface liquid (ASL) appears to be slightly acidic (pH 6.9), and acidity increases in CF. Cells in the submucosal glands secrete mucus, and secretion of (primarily) HCO3 and (some) Cl along with Na through tight junctions (to maintain electroneutrality) and fluid sweeps the mucus out of the glands. Permeability of tight junctions between submucosal gland cells is critical for the glands' ability to secrete ions. In contrast, surface cells seem to have tight junctions that are impermeant to ions, and this assures that these cells operate only to absorb Na (through ENaC), Cl (through CFTR) and water. According to these proposals, the absence of CFTR in CF will cause: (i) decreased HCO3, Na and water secretion by submucosal gland cells (leading to mucus clogging); (ii) increased acidity of ASL (due to reduced HCO3 secretion); (iii) thicker and stickier mucus (due to reversal of fluid secretion to absorption and increased acidity of ASL). According to these hypotheses, it is crucial to determine mechanisms of H and HCO3 transport across the membranes and tight junctions of WT and CF submucosal gland and surface epithelial cells, including the potential role of mucus secretion to the acid-base balance of the ASL. The specific aims of this proposal are: (i) Use fluorescence microscopic imaging, immunomicroscopy and western blots to identify mechanisms for HCO3 and H transport across apical and basolateral membranes of submucosal gland and surface cells of WT and CF airway monolayers. (ii) Using genetic targeting of pH-sensitive dyes and imaging microscopy, measure pH (and regulation) in mucus granules and at the lumenal aspect of WT and CF airway epithelia. (iii) Using isotopic methods, measure secretion of mucus and of constitutive and regulated Golgi-derived vesicles and correlate to H/HCO3 transport. (iv) Using electrophysiological, isotopic flux and immunomicroscopic methods, determine the permeability/conductance properties of the tight junctions.

描述（申请人摘要）：气道上皮细胞的酸碱转运 由于对粘液的影响，细胞可能在囊性纤维化（CF）中发挥关键作用 水化和流变学、纤毛运输、细菌结合和抗微生物 因素气道表面液体（ASL）似乎呈弱酸性（pH 6.9）， CF中的酸度增加。粘膜下腺体中的细胞分泌粘液， （主要）HCO 3和（一些）Cl沿着Na的分泌， 连接（以维持电中性）和流体清扫粘液出的 腺体粘膜下腺细胞间紧密连接的通透性 对腺体分泌离子的能力至关重要相反，表面细胞 似乎有紧密的连接，是不渗透的离子，这保证了， 这些细胞只吸收Na（通过ENaC），Cl（通过CFTR）和 水根据这些建议，CF中没有CFTR将导致： 减少粘膜下腺细胞的HCO 3、Na和水分泌（导致 （ii）ASL的酸度增加（由于HCO 3分泌减少）; (iii)更粘稠的粘液（由于液体分泌逆转， ASL的吸收和增加的酸度）。根据这些假设， 确定H和HCO 3跨膜运输的机制至关重要 WT和CF粘膜下腺体和表面上皮细胞的紧密连接， 包括粘液分泌对胃粘膜酸碱平衡的潜在作用。 美国手语本建议的具体目标是：（一）使用荧光显微镜 成像，免疫显微镜和蛋白质印迹，以确定HCO 3的机制， H穿过粘膜下腺体的顶膜和基底外侧膜的转运， WT和CF气道单层的表面细胞。(ii)利用基因靶向 pH敏感染料和成像显微镜，测量粘液中的pH值（和调节） 颗粒和WT和CF气道上皮细胞的管腔方面。(iii)使用 同位素方法，测量粘液分泌以及组成性和调节性 高尔基体衍生的囊泡和相关的H/HCO 3运输。(iv)使用 电生理、同位素通量和免疫显微镜方法，测定 紧密连接的渗透性/传导性。

项目成果

Pseudomonas aeruginosa Inhibition of Flagellin-activated NF-kappaB and interleukin-8 by human airway epithelial cells.

铜绿假单胞菌通过人气道上皮细胞抑制鞭毛蛋白激活的 NF-kappaB 和白细胞介素 8。

• DOI: 10.1128/iai.01355-08
• 发表时间: 2009
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Pena,Jose;Fu,Zhu;Schwarzer,Christian;Machen,TerryE
• 通讯作者: Machen,TerryE

Modulation of cytosolic Ca(2+) concentration in airway epithelial cells by Pseudomonas aeruginosa.

铜绿假单胞菌对气道上皮细胞胞浆 Ca(2) 浓度的调节。

• DOI: 10.1128/iai.70.11.6399-6408.2002
• 发表时间: 2002
• 期刊: Infection and immunity
• 影响因子: 3.1
• 作者: Jacob,Tobias;Lee,RebeccaJ;Engel,JoanneN;Machen,TerryE
• 通讯作者: Machen,TerryE

Organelle redox of CF and CFTR-corrected airway epithelia.

CF 和 CFTR 校正的气道上皮细胞的细胞器氧化还原。

• DOI: 10.1016/j.freeradbiomed.2007.04.015
• 发表时间: 2007
• 期刊: Free radical biology & medicine
• 影响因子: 7.4
• 作者: Schwarzer,Christian;Illek,Beate;Suh,JungH;Remington,SJames;Fischer,Horst;Machen,TerryE
• 通讯作者: Machen,TerryE