PERTICIPATION OF INTRACELLULAR SIGNAL TRANSDUCTION PATHWAY AND RE-PERFUSION LUNG INJURY AFTER LUNG TRANSPLANTATION

细胞内信号转导通路与肺移植后再灌注肺损伤的关系

• 批准号: 13470266
• 负责人: TANITA Tatsuo
• 金额: $ 9.22万
• 依托单位: IWATE MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470266/
• 关键词: LUNG TRANSPLANTATITON; LUNG INJURY; PULMONARY VASCULAR ENDOTHELIAL CELLS; INTRACELLUL SIGNAL TRANSDUCTION; PROTEIN KINASE C; ACTIN POLYMERIZATION; TRANSENDOTHELIAL ELECTRICAL RESISTANCE

We showed that neutrophils-induced lung injury was induced by reactive oxygen species generated via xanthine oxidase(XO) following adhesion of neutrophils on the endothelial cells; Usually, XO does not exist in the neutrophils but endothelial cells. Therefore we speculate if this kind of lung injury is induced by activation of endothelial cells themselves. In the experiments of isolated perfused rat lungs, whether intracellular signal transduction systems were involved in the injury of the pulmonary vascular endothelial cells caused by activated neutrophils, we investigated using a protein kinase C antagonist. We showed that PKC antagonist ameliorated the pulmonary vascular injury caused by activated neutrophils in a dose dependent manner. Besides, we showed that the pulmonary vascular permeability was increased by a PKC agonist without neutrophils. On the other hand, in cell culture models, it was impossible to measure protein permeability for cell culture sheets, we measured trans-en … More dothelial electrical resistance (TER) across the cultured pulmonary vascular endothelial cell sheets. The dominanat effect of hyperosmolar exposure was an increase in the TER, indicating the induction of barrier strengthening. Hyperosmolar exposure also increased activity of focal adhesion kinase and E-cadherin accumulation at the cell periphery. Concomitantly, the density of actin filaments increased markedly. PMA (phobor myristate acetate), an agonist of Protein kinase C, decreased the TER, indicating a de-polymerization of actin filaments. On the other hand, cytochalasin D, de-polymerize agents for actin filaments, also decreased TER, indicating participation of actin de-polymerization. The actin response to hyperosmolar exposure was enhanced although the response in TER was unaffected, indicating that the endothelial barrier enhancement occurred independently actin. However, in monolayers expressing a kinase-dependent mutant of focal adhesion kinase, the hyperosmolarity-induced increase in activity of focal adhesion kinase and peripheral E-cadherin enhancement were blocked and the induced increase of TER was markedly blunted. These findings indicate that EC exposed to hyperosmolar challenge, the involvement of focal adhesion kinase was critical in establishing barrier strengthening. Less

我们发现，中性粒细胞引起的肺损伤是由中性粒细胞与内皮细胞黏附后产生的黄嘌呤氧化酶(XO)产生的活性氧引起的；通常中性粒细胞中不存在XO，而是内皮细胞中存在XO。因此，我们推测这种肺损伤是否是由内皮细胞本身激活引起的。在大鼠离体肺灌流实验中，我们用蛋白激酶C拮抗剂研究了细胞内信号转导系统是否参与了中性粒细胞活化对肺血管内皮细胞的损伤。我们发现PKC拮抗剂以剂量依赖的方式改善中性粒细胞激活所致的肺血管损伤。此外，我们还发现不含中性粒细胞的PKC激动剂可增加肺血管通透性。另一方面，在细胞培养模型中，不可能测量细胞培养片的蛋白质渗透性，我们测量了反式…培养的肺血管内皮细胞膜有更多的内皮电阻(TER)。高渗暴露的主要效应是TER增加，提示屏障增强。高渗暴露也增加了粘着斑激酶的活性和E-钙粘附素在细胞外围的积聚。肌动蛋白细丝密度明显增加。蛋白激酶C的激动剂PMA(肉豆蔻酸酯)降低了TER，表明肌动蛋白细丝解聚。另一方面，肌动蛋白细丝的解聚剂细胞松弛素D也降低了TER，表明参与了肌动蛋白解聚。肌动蛋白对高渗暴露的反应增强，而TER的反应不受影响，说明内皮细胞屏障的增强是独立发生的。然而，在表达粘着斑激酶突变体的单层细胞中，高渗诱导的粘着斑激酶活性的增加和外周E-钙粘附素的增强被阻断，TER的增加被明显钝化。这些结果表明，EC暴露在高渗刺激下，粘着斑激酶的参与在建立屏障强化中起着关键作用。较少

项目成果

S K Quadri, M Bhattacharjee, K Parthasarathi, et al.: "Endothelial Barrier Strengthening by Activation of Focal Adhesion Kinase."J Biol Chem. 278. 13342-13349 (2003)

S K Quadri、M Bhattacharjee、K Parthasarathi 等人：“通过激活局部粘附激酶来增强内皮屏障。”J Biol Chem。

Tanita T, Song C, Kubo H, Hoshikawa Y, Chida M, Suzuki S, Ono S, Fujimura S.: "Superoxide anion mediates pulmonary vascular permeability caused by neutrophils in cardiopulmonary bypass."Surg Today. 29. 755-761 (1999)

Tanita T、Song C、Kubo H、Hoshikawa Y、Chida M、Suzuki S、Ono S、Fujimura S.：“超氧阴离子介导体外循环中中性粒细胞引起的肺血管通透性。”Surg Today。

Sadiqa K.Quadri, Mrinal: "Bhattacharjee, Kaushik Parthasarathi, Tatsuo Tanita, Jahar Bhattacharya. Endothelial Barrier Strengthening by Activation of Focal Adhesion Kinase."J Biol Chem. 278. 13342-13349 (2003)

Sadiqa K.Quadri，Mrinal：“Bhattacharjee、Kaushik Parthasarathi、Tatsuo Tanita、Jahar Bhattacharya。通过激活局部粘附激酶来增强内皮屏障。”《生物化学》杂志。

Hoshikawa Y, Ono S, et al.: "Generation of oxidative stress contributes to the development of pulmonary hypertension induced by hypoxia."J Appl Physiol. 90. 1299-1306 (2001)

Hoshikawa Y、Ono S 等人：“氧化应激的产生有助于缺氧引起的肺动脉高压的发生。”J Appl Physiol。

Quadri SK: "Endothelial barrier strengthening by activation of focal adhesion kinase"J Biol Chem. 278・15. 13342-13349 (2003)

Quadri SK：“通过粘着斑激酶的激活增强内皮屏障”J Biol Chem. 278・15。