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The effect of cytokines on leukocyte kinetics in rat pulmonary microcirculation.

细胞因子对大鼠肺微循环白细胞动力学的影响。

基本信息

批准号：
07671449
负责人：
KITAMURA Michihiko
金额：
$ 1.34万
依托单位：
Akita University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671449/
关键词：
lung injury neutrophil hydrogen peroxide cytokine 活性酸素肺障害肺循環

项目摘要

Although, it was reported that cytokines and the leukocyte play major roles in acute lung injury, the effect of cytokines on the leukocyte kinetics in the pulmonary microcirculation is not fully understood. To investigate the role of leukocytes in the rat acute lung injury model, we developed two novel methods to detect hydrogen peroxide in the rat and rabbit pulmonary microcirculation. We applied the fluorescent dye, 2', 7'-dichlorofluorescin diacetate (DCFH-DA), which has been used to measure hydrogen peroxide generated from isolated neutrophils by the flowcytometry, to a live animal. We also applied the cerium technique, which has been used to detect hydrogen peroxide generated from an isolated neutrophil by the electron microscopy, to a live animal. The acute lung injury rat model was made by a continuous infusion of endotoxin at a rate of 5 mg/kg/hr. Using these techniques, we detected and quantified the hydrogen peroxide production in the neutrophil adhered to the pulmonary capillary endothelium. We also found that this hydrogen peroxide diffuses into the endothelial cell. The rat lung infused with endotoxin was immunostained with the rabbit polyclonal antibody against rat TNFalpha. The alveolar wall of the rat was positively stained with that. We also measured TNFa contents in the rat lung infused with endotoxin by ELISA method. The contents of TNFalpha in the rat lung infused with endotoxin was five times higher than that in the control rat lung. In this project, we demonstrated roles of neutrophil and TNFalpha in the pulmonary microcirculation of the acute lung injury animal model.

虽然有报道称细胞因子和白细胞在急性肺损伤中起主要作用，但细胞因子对肺微循环中白细胞动力学的影响尚未完全了解。为了探讨白细胞在大鼠急性肺损伤模型中的作用，我们建立了两种检测大鼠和兔肺微循环中过氧化氢的新方法。我们将荧光染料2 '，7'-二氯荧光素二乙酸酯（DCFH-DA）应用于活体动物，该染料已用于通过流式细胞术测量分离的中性粒细胞产生的过氧化氢。我们还应用了铈技术，这已被用来检测过氧化氢产生的一个孤立的中性粒细胞的电子显微镜，一个活的动物。以5 mg/kg/hr的速率持续输注内毒素造成急性肺损伤大鼠模型，应用上述技术检测并定量肺毛细血管内皮细胞粘附的中性粒细胞产生的过氧化氢。我们还发现这种过氧化氢扩散到内皮细胞中。用兔抗大鼠TNF α多克隆抗体对内毒素灌注的大鼠肺进行免疫染色。大鼠肺泡壁呈阳性染色。用ELISA法测定内毒素灌注大鼠肺组织中TNF α的含量。内毒素组大鼠肺组织TNF α含量较对照组高5倍。本课题研究了中性粒细胞和肿瘤坏死因子α在急性肺损伤动物模型肺微循环中的作用。