Suzuki Y, Aoki T, Suzuki K, Miyata A, Nishio K, Tsumura K, Takasugi T, Mori M, Suematsu M, Yamaguchi K
Department of Internal Medicine, Kitasato Institute Hospital, Tokyo, Japan.
Nihon Kyobu Shikkan Gakkai Zasshi. 1997 Feb;35(2):137-43.
To study the dynamic interaction between blood cells and endothelial cells in the pulmonary microcirculation, we developed a method for observing leukocytes and erythrocytes in the microcirculation of perfused rat lungs by confocal laser scanning microscopy. We examined the behavior of leukocytes in the microcirculation of rat lungs exposed to hyperoxia, because oxygen toxicity is known to be associated with leukocyte infiltration and endothelial cell damage. Rats were divided into two groups: control (21% O2) and hyperoxia (90% O2 for 48 hours). Lungs were perfused with Krebs-Henseleit solution equilibrated with 21% O2 and 5% CO2 through an artificial lung. Leukocytes stained with carboxyfluorescein diacetate succinimidyl ester and erythrocytes stained with fluorescein isothiocyanate were added to the perfusate, and images of the cells were observed and recorded with a confocal laser scanning microscope and a high-speed video camera. The mean velocities of erythrocytes (Vr) in arterioles, capillaries, and venules of the control group were 1.52, 0.50 and 1.61 mm/sec, respectively. Also in the control group, the velocities of the leukocytes were divided by the mean velocities of the erythrocytes in the same arterioles, capillaries and venules (Vw/Vr) and the results were 0.96, 0.97, and 0.98, respectively. The Vw/Vr values for arterioles in the hyperoxia group were not significantly different from those in the control group, but the Vw/Vr values for capillaries and venules were 27% and 37% lower than their respective control values. Leukocyte sequestration was seen mainly in capillaries in the hyperoxic group. These results suggest that an increase in adhesion in capillaries and venules, such as that caused by adhesion molecules, might play a key role in the behavior of leukocytes in the pulmonary microcirculation during hyperoxia.
为了研究肺微循环中血细胞与内皮细胞之间的动态相互作用，我们开发了一种通过共聚焦激光扫描显微镜观察灌注大鼠肺微循环中白细胞和红细胞的方法。我们研究了暴露于高氧环境下大鼠肺微循环中白细胞的行为，因为已知氧中毒与白细胞浸润和内皮细胞损伤有关。大鼠分为两组：对照组（21%氧气）和高氧组（90%氧气，持续48小时）。通过人工肺用含21%氧气和5%二氧化碳平衡的克雷布斯 - 亨泽莱特溶液灌注肺。将用羧基荧光素二乙酸琥珀酰亚胺酯染色的白细胞和用异硫氰酸荧光素染色的红细胞加入灌注液中，并用共聚焦激光扫描显微镜和高速摄像机观察并记录细胞图像。对照组小动脉、毛细血管和小静脉中红细胞的平均速度（Vr）分别为1.52、0.50和1.61毫米/秒。同样在对照组中，将小动脉、毛细血管和小静脉中白细胞的速度除以同一血管中红细胞的平均速度（Vw/Vr），结果分别为0.96、0.97和0.98。高氧组小动脉的Vw/Vr值与对照组无显著差异，但毛细血管和小静脉的Vw/Vr值分别比各自对照组值低27%和37%。高氧组中白细胞滞留主要见于毛细血管。这些结果表明，毛细血管和小静脉中黏附增加，如由黏附分子引起的黏附增加，可能在高氧期间肺微循环中白细胞的行为中起关键作用。
