APOPTOSIS IN ACUTE LUNG INJURY

急性肺损伤中的细胞凋亡

基本信息

批准号：
10470322
负责人：
HASHIMOTO Satoru
金额：
$ 5.06万
依托单位：
KYOTO PREFECTURAL UNIVERSITY OF MEDICINE
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B).
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10470322/
关键词：
Acute lung injury Apoptosis ARDS Fas FasL

项目摘要

Accumulation and activation of inflammatory cells in the lung characterize the acute respiratory distress syndrome (ARDS). However, the precise mechanism for lung epithelial and endothelial cell damage remains unknown. Based on evidence that rapid apoptosis caused by CD8^+ cytolytic T cells can induce pathological cell death, we hypothesized that this mechanism may also participate in the acute lung injury. To determizae the possible contribution of apoptosis in the pathogenesis of acute lung injury (ALI), we investigated Fas antigen (Fas), Fas ligand (FasL), perforin, granzyme A, and granzyme B expressions in septic ARDS patients, and in a murine model of ALI after intratracheal instillation of Escherichia coli lipopolysaccharide (LPS) into the left lung. Quantitative PCR analysis revealed that the mRNAs for several apoptosis molecules were highly upregulated in the acute phase of ARDS following sepsis. The level of soluble FasL in the BALF increased only in the acute ARDS patients. W … More hile, in a murine LPS model, expressions of the pro-apoptosis molecules' mRNA were dose-dependently upregulated, with maximal expression in the early phase in the instilled lung and most apparent one day after LPS-instillation. Negligible mRNA expression of pro-apoptosis molecules was observed in non-instilled lungs. The terminal deoxynucleotidyl-transferase mediated dUTP biotin nick end labeling (TUNEL) demonstrated positive signals in neutrophils and macrophages as well as in alveolar wall cells of the instilled lung one day after the LPS-instillation. Immunohistochemistry demonstrated that Fas was upregulated in alveolar and inflammatory cells and FasL-positive inflammatory cells migrated into the air spaces in the LPS-instilled lung. Intratracheal administration of P2 antibody, which is an anti-Fas blocking antibody, attenuated the lung injury after 30 μg LPS-instillation without attenuating mRNA expressions of pro-apoptosis molecules and neutrophil accumulation in the lung. These results indicate that Fas/FasL system could be important in the pathogenesis of ALI, and proper regulation of FasL/Fas system might be important for potential ARDS treatment. Less

肺部炎症细胞的积累和激活是急性呼吸窘迫综合征 (ARDS) 的特征，然而，基于 CD8^+ 溶细胞 T 细胞引起的快速细胞凋亡的确切机制仍不清楚。病理性细胞死亡，我们发现这种机制也可能参与急性肺损伤。为了确定细胞凋亡在急性肺损伤（ALI）发病机制中的可能作用，我们研究了 Fas 抗原（Fas），脓毒症 ARDS 患者以及左肺气管内滴注大肠杆菌脂多糖 (LPS) 后的 ALI 小鼠模型中 Fas 配体 (FasL)、穿孔素、颗粒酶 A 和颗粒酶 B 的表达定量 PCR 分析显示，脓毒症后 ARDS 的急性期，多种凋亡分子高度上调 BALF 中可溶性 FasL 水平。仅在急性 ARDS 患者中增加，而在小鼠 LPS 模型中，促凋亡分子 mRNA 的表达呈剂量依赖性上调，在注入肺的早期阶段表达量最大，一天最明显。在未滴注的肺中观察到LPS滴注后，末端脱氧核苷酸转移酶介导的dUTP生物素的mRNA表达可忽略不计。 LPS 滴注一天后，切口末端标记 (TUNEL) 显示中性粒细胞和巨噬细胞以及注入肺的肺泡壁细胞中出现阳性信号。免疫组织化学显示肺泡和炎症细胞中的 Fas 上调，并且 FasL 阳性炎症细胞迁移。气管内注射 P2 抗体，这是一种抗 Fas 阻断抗体， 30 μg LPS 滴注后减轻了肺损伤，但不减弱促凋亡分子的 mRNA 表达和肺中中性粒细胞的积累。这些结果表明 Fas/FasL 系统在 ALI 的发病机制以及 FasL/Fas 的适当调节中可能很重要。系统对于潜在的 ARDS 治疗可能很重要。