Dynamic Analysis of Hepatic Microcirculation and Hepatic Function by In vivo-Fluorescence Microscopy -Application to the Study of Pathogenesis and Treatment of Liver Diseases.

采用活体荧光显微镜观察肝脏微循环（超过2000X），阐明酒精、药物和病毒致肝损伤过程中细胞功能的动态变化。对血流量进行连续定量测量。红细胞和白细胞的正弦速度。血氧饱和度等等。肝小叶微循环表现为：中心周围（PC）区血流速度快于门静脉周围（PP）区血流速度；血氧饱和度从PP区向PC区呈梯度（PP>PC）。此外，我们还评估了乙醇对肝脏微循环的影响，以阐明微循环障碍在酒精性肝损伤机制中的参与。低剂量酒精增加了正弦血流量和正弦血氧合，但部分PC区正弦血氧合下降，对酒精有异质反应。相反，高剂量酒精显著降低了正脉血的氧合，特别是在PC区，导致缺氧区数量增加。此外，使用该系统的荧光素钠研究显示肝细胞向胆管的排泄受损，特别是在PC区。这些数据表明，高剂量酒精产生小叶内分流和局部缺氧，导致肝功能障碍。在灌注的肝脏中，酒精通过涉及细胞内钙的机制引起血管血管前区收缩。总之，本科学研究所建立的方法不仅对阐明肝病的病理生理，而且对肝病的治疗和预防都是一个有用的工具。

The in vivo fluorescence microscopy was developed to visualize the microcirculation (over 2000X) of the liver to elucidate the dynamic change of cellular function in the process of hepatic injury caused by alcohol, drug and virus. The continuous and quantitative measurement was performed for the sinusoidal blood flow. Sinusoidal velocity of red and white blood cells. Oxygen saturation of the sinusoidal blood and so on. The microcirculation of the liver lobule was characterized as follows: The sinusoidal blood flow was faster in the pericentral(PC) region than in periportal(PP) region; there was a gradient of the oxygen saturation of sinusoidal blood from PP to PC area(PP>PC). In addition, the effect of ethanol on the hepatic microcirculation was evaluated in order to clarify the involvement of microcirculatory disturbance in the mechanism of alcoholic liver injury. Low dose alcohol raised both the sinusoidal blood flow and oxygenation of sinusoidal blood, yet the oxygenation of sinusoidal blood fell in some sinusoids of PC region, giving a heterogenic response to alcohol. In contrast, high dose alcohol markedly reduced the oxygenation of sinusoidal blood particularly in PC region, resulting in an increased number of hypoxic regions. Furthermore, sodium fluorescein study using this system showed an impairment in the excretion from hepato-cytes to bile canaliculi particularly in PC area. These data indicate that high dose alcohol produces intralobular shunting and local hypoxia, leading to hepatic dys-function. In the perfused liver, it was demonstrated that alcohol caused a vaso-constriction at presinusoidal region via mechanism involving intracellular calcium. In conclusion, the methodology developed in this scientific research is a useful tool not only for the ellucidation of pathophysiology of the liver disease, but for the treatment and prevention of liver disease.