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A study for pulmonary cell preservation and lung transplantation on functions of alveolar epithelial ion transport

肺细胞保存及肺移植对肺泡上皮离子转运功能的影响研究

基本信息

批准号：
08457342
负责人：
TANITA Tatsuo
金额：
$ 4.54万
依托单位：
Tohoku University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08457342/
关键词：
Chronic pulmonary emphysema alveolar epithelial cells Ion transport Pulmonary cell preservation Lung injury Cell culture Lung preservation medium Steroid 肺障害肺胞II型上皮細胞 enzymatic tissue digestion法 difference adherence法 Na+-K+-ATPase

项目摘要

In patients of chronic pulmonary emphysema or pulmonary fibrosis, pulmonary epithelial cells were irreversibly damaged. These phenomena, combined with recent advancing of age of population, are one of the important reasons for an increase in number of chronic respiratory failure. Therefore, we started to study whether transplantation of pulmonary type 2 cells are feasible for a therapy of impairment of pulmonary epithelial functions.In 1996, we established a method of separation and culture for pulmonary type 2 cells, and a method of measurement of activity of Na^+-K^+-ATPase, which is the main function of pulmonary type 2 cells.ln 1997, we tried to establish a more effective separation method of pulmonary type 2 cells using a cell sorter. Although, this new method has not been established yet, we could separate pulmonary type 2 cells with purity of more than 80 %.Then we tried to establish a method for keeping viability of pulmonary type 2 cells by studying pulmonary cellular preservation. We compared pulmonary preservation mediums EP4 solution, established by our institute, and Euro-Collins solution (EC), usual preservation medium. When we measured alveolar epithelial functions using Alveolar Fluid Clearance (AFC), EP4 maintained alveolar epithelial functions but EC could not maintain those.In lung injury models, we showed that cyclooxygenase inhibitor prevented the pulmonary vascular permeability. Therefore, we tested steroid for function of alveolar epithelial cells. In this experiment, gluco-corticoid increased AFC.This is a new evidence that steroid acts not only as an anti-inflammatory agent but to protect living body from lung injury by pumping up alveolar fluid to lung interstitial space.

在慢性肺气肿或肺纤维化患者中，肺上皮细胞受到不可逆的损伤。这些现象，加上人口年龄的增长，是慢性呼吸衰竭数量增加的重要原因之一。因此，我们开始研究肺2型细胞移植治疗肺上皮功能障碍的可行性，1996年，我们建立了肺2型细胞的分离培养方法和肺2型细胞主要功能Na^+-K^+-ATP酶活性的测定方法，1997年，我们尝试使用细胞分选仪建立更有效的肺2型细胞分离方法。虽然这种新的方法尚未建立，但我们可以分离出纯度在80%以上的肺2型细胞，因此我们通过对肺细胞保存的研究，试图建立一种保持肺2型细胞活力的方法。比较了我所研制的EP 4液和常用的保存液Euro-Collins液（EC液）的保存效果。用肺泡液体清除率（AFC）测定肺泡上皮功能，发现EP 4能维持肺泡上皮功能，而EC不能维持肺泡上皮功能。因此，我们测试了类固醇对肺泡上皮细胞的功能。本实验中糖皮质激素可增加AFC，这为糖皮质激素不仅具有抗炎作用，而且通过将肺泡液泵入肺间质而保护机体免受肺损伤提供了新的证据。