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The evaluation of organ injuries due to surgical stress using green fluorescent protein induced rat.

使用绿色荧光蛋白诱导大鼠评估手术应激引起的器官损伤。

基本信息

批准号：
15591488
负责人：
SATO Yukio
金额：
$ 1.92万
依托单位：
JICHI MEDICAL SCHOOL
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591488/
关键词：
Green flurescent protein Polymorphonuclear leukocyte Acute lung injury Nitric oxide Phoshpodiesterase

项目摘要

Activated PMN sequester in pulmonary microvessels and play an important role in acute lung injury. However, the site of PMN sequestration is still controversial, as in vivo real time observation of PMN in pulmonary microvessels has been difficult.We developed a new method using GFP transgenic rat. Rats were anesthetized, ventilated and catheters were placed in the superior vena cava and the aortic root for the administration of agents and the monitoring of blood pressure. The left anterolateral chest wall was opened, the left lung was stabilized and PMN with GFP was viewed under a fluorescent microscope. The image was obtained with CCD camera and recorded in a hard disk recorder.Control group (n=4) received the injection of saline. Lipopolysaccaride(LPS) treated group received the injection of LPS (1mg/kg) from Escherichia coli. PMN count decreased in LPS treated group (2610±580 to 1180±220/μl, p<0.05), while PMN count did not change in control (2490±700 to 3320±960/μl). Systolic blood pressure and heart rate did not change in both groups (LPS;121±9 to 123±8mmHg, 319±19 to 295±20/min, control;133±27 to 135±30 mmHg,322±22 to 292±22/min). LPS treatment decreased PMN velocity passing through capillaries from 421±86 to 51±21 μm/sec and increased sequestered PMN in capillaries. The population of rolling leukocytes in postcapillary venule was less than 10 % and LPS treatment did not increase rolling of PMN. Endotoxemia decreased PMN velocity passing through capillaries and increased PMN sequestration in capillaries, but did not change rolling of leukocyte in postcapillary venule. We further evaluated the effect of nitric oxide and phosphodiesterase type 4 inhibitor on PMN sequestration in pulmonary microcirculation.

活化的PMN在肺微血管中隔离并在急性肺损伤中起重要作用。然而，PMN隔离的位置仍然存在争议，因为在肺微血管中实时观察PMN一直很困难。我们开发了一种利用转基因大鼠的新方法。大鼠麻醉、通气，在上腔静脉和主动脉根部放置导管给药并监测血压。打开左前外侧胸壁，稳定左肺，荧光显微镜下观察含GFP的PMN。图像由CCD摄像机采集，并记录在硬盘记录仪中。对照组（n=4）给予生理盐水注射。脂多糖（LPS）处理组给予大肠杆菌LPS （1mg/kg）注射。LPS处理组PMN计数下降（2610±580 ~ 1180±220/μl, p<0.05），对照组PMN计数无变化（2490±700 ~ 3320±960/μl）。两组的收缩压和心率均无变化（LPS组；121±9 ~ 123±8mmHg, 319±19 ~ 295±20/min，对照组；133±27 ~ 135±30 mmHg,322±22 ~ 292±22/min）。LPS处理降低了PMN通过毛细血管的速度，从421±86 μm/秒降低到51±21 μm/秒，增加了毛细血管中被隔离的PMN。毛细血管后小静脉中滚动白细胞的数量少于10%，LPS处理没有增加PMN的滚动。内毒素血症降低了PMN通过毛细血管的速度，增加了PMN在毛细血管的固存，但没有改变白细胞在毛细血管后小静脉的滚动。我们进一步评估了一氧化氮和磷酸二酯酶4型抑制剂对肺微循环中PMN封存的影响。