1996 Fiscal Year Final Research Report Summary
The effect of cytokines on leukocyte kinetics in rat pulmonary microcirculation.
| Project/Area Number |
07671449
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | Akita University |
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Principal Investigator |
KITAMURA Michihiko Akita University School of Medicine Associate Professor, 医学部, 助教授 (10153131)
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| Co-Investigator(Kenkyū-buntansha) |
MINAMIYA Yoshihiro Akita University School of Medicine Assistant Professor, 医学部, 講師 (30239321)
IZUMI Keiichi Akita University School of Medicine Assistant Professor, 医学部, 講師 (60176237)
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| Project Period (FY) |
1995 – 1996
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| Keywords | lung injury / neutrophil / hydrogen peroxide / cytokine |
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| Research Abstract |
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.
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