Role of Alveolar Type II Epithelial Cells in the Repairing Process after Lung Injury; Stimulating Factors for Cell Proliferation and Ion Transport

肺泡 II 型上皮细胞在肺损伤后修复过程中的作用；

• 批准号: 62570705
• 负责人: SUGAHARA Kazuhiro
• 金额: $ 1.66万
• 依托单位: KUMAMOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62570705/
• 关键词: Alveolar type II epithelial cells; Surface active material; Lung injury; Repair and recovery; Growth factor; 人体材料

1. Isolation and culture of human alveolar type II cells from resected human lungs: Specimens were obtained from ll patients undergoing lobectomy or pneumonectomy for primary tumors. Alveolar type II cells were isolated by a modification of the method for isolating rat type II cells with elastase and density gradients. The yield of type II cells was 2.7 x10^6 cells/g of tissue weigh. the percentage of type ii cells as determined by the papanicolaou stain after one day in culture was 71.0 3.4 % in 8 of ll isolations. These human cells have more numerous and smaller granules than do isolated rat type II cells. EM of the cells showed characteristic surface microvilli and intracytoplasmic lamellar bodies, of which some were multicentric. Examining the cytoskeleton structure by an avidin-biotin stain, they have fine intermediate filaments such as actin and keratin. We measured the secretion of pulmonary surfactant apoprotein from human type II cells over 3-6 hours after one day in culture. … More Apoprotein was secreted about 33.1 ng/10^6 cells in 3 hrs, and 35.3 ng/10^6 cells in 6 hrs. This secretion was stimulated almost 2-fold by ADP 10^<-3>M.2. Functions of alveolar type II cells: (1) effect of silica in vitro; Alveolar type II cells appear hypertrophic and hyperlasic in silica-treated lungs, we investigated type II cell proliferations, measuring tritiated thymidine incorporation. Co-culturing type II cells with alveolar macrophages stimulated by silica, using transwell membrane, thymidine incorporation was inhibited to 46.9 % of the control. (2) in vivo culture; Altough the cells are necessarily damaged and lose surface receptors to some extent by the isolation procedure and conditions in vitro, we develop a new system of in vivo culture, which isolated cells are again implanted into the peritoneum or subcutaneous region of another rat, using diffusion chambers. The cells in vivo culture more proliferated and had more active ion transport. (3) effect of extracellular matrix; Fibronectin mediates alveolar type II cells adherence in vitro. Monolayers on fibronection coated filter have higher pothtial difference and resistance. Fluorescence microscopy of type II cells on fibronectin showed a marked accumulation of actin and keratin bundles in their periphery. Fibronectin is an important functional component of the extracellular matrix that may support alveolar type II cells during the alveolar re-epithelization after lung injury.3. Function of tracheal epithelium: The tracheal epithelium have some similar functions to those of alveolar type II cells, such as ion transport and cell proliferation. Less

1.从切除的人肺中分离和培养人肺泡II型细胞：从因原发性肿瘤而进行肺叶切除术或肺切除术的11名患者中获得标本。肺泡II型细胞分离的方法与弹性蛋白酶和密度梯度分离大鼠II型细胞的修改。II型细胞的产量为2.7 × 10^6个细胞/g组织重量。在培养一天后通过巴氏染色测定的II型细胞的百分比在8个L1分离物中为71.0 ± 3.4%。这些人类细胞比分离的大鼠II型细胞具有更多和更小的颗粒。电镜下可见特征性的表面微绒毛和胞浆内板层体，部分板层体为多中心结构。用抗生物素蛋白-生物素染色法检查细胞骨架结构，它们具有细的中间丝，如肌动蛋白和角蛋白。我们测量了培养一天后3-6小时内人II型细胞的肺表面活性物质脱辅基蛋白的分泌。 ...更多信息 载脂蛋白在3小时内分泌约33.1 ng/10^6个细胞，在6小时内分泌约35.3 ng/10^6个细胞。ADP 10 μ M几乎可刺激该分泌2倍<-3>。肺泡II型细胞的功能：（1）二氧化硅的体外作用：肺泡II型细胞在二氧化硅处理的肺中出现肥大和增生，我们研究了II型细胞的增殖，测量氚化胸苷掺入。将Ⅱ型细胞与经二氧化硅刺激的肺泡巨噬细胞共培养，使用transwell膜，胸苷掺入被抑制至对照的46.9%。(2)体内培养;尽管细胞在体外的分离过程和条件下必然会受到一定程度的损伤并失去表面受体，但我们开发了一种新的体内培养系统，将分离的细胞再次植入另一只大鼠的腹膜或皮下区域，使用扩散室。体内培养的细胞增殖更快，离子转运更活跃。(3)细胞外基质的作用;纤连蛋白介导肺泡II型细胞粘附的体外研究。纤维连接蛋白膜上的单分子膜具有较高的电位差和电阻。荧光显微镜下的II型细胞纤连蛋白显示了显着积累的肌动蛋白和角蛋白束在其周边。纤维连接蛋白是细胞外基质的重要功能成分，在肺损伤后肺泡上皮再形成过程中可能支持肺泡II型细胞.气管上皮的功能：气管上皮具有与肺泡Ⅱ型细胞相似的功能，如离子转运和细胞增殖。少

项目成果

須加原一博: 日本界面医学会雑誌. 19. 148-151 (1988)

Kazuhiro Sukahara：日本界面医学学会杂志 19. 148-151 (1988)。

Kazuhiro SUGAHARA; Taketoshi KIYOTA; Tomoko BABA et al: "Ion transport by alveolar epithelial cells" KOKYU TO JUNKAN. 36. 609-614 (1988)

菅原一宏；

須加原一博: 呼吸と循環. 36. 609-614 (1988)

Kazuhiro Sukahara：呼吸和循环。36。609-614（1988）

Taketoshi KIYOTA.: Japanese Anasthesia Journals' Review. 3. (1988)

Taketoshi KIYOTA.：日本麻醉期刊评论。

Taketoshi KIYOTA: Japanese Anaesthesia Journals' Review. 3. (1988)

Taketoshi KIYOTA：日本麻醉期刊评论。