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

为了阐明酒精、药物和病毒引起的肝损伤过程中细胞功能的动态变化，建立了活体荧光显微镜来显示肝脏微循环(大于2000倍)。对正弦血流量进行连续定量测量。红细胞和白细胞的正弦运动速度。正弦血氧饱和度等。肝小叶微循环的特点是：中心周围(PC)区的肝窦血流速度快于门脉周围(PP)区，肝门周围区(PP&GT；PC)的肝小叶血氧饱和度存在梯度。此外，还观察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作用。低剂量酒精使正弦血流量和血氧饱和度均升高，但PC区部分正弦血氧饱和度下降，对酒精有异质性反应。相反，高剂量酒精显著降低血窦血液的氧合，特别是PC区，导致缺氧区的数量增加。此外，使用该系统的荧光素钠研究表明，肝细胞向胆小管的排泄受到损害，尤其是PC区。这些数据表明，高剂量酒精引起小叶内分流和局部缺氧，导致肝功能障碍。在灌流的肝脏中，证实了酒精引起窦前区血管收缩的机制与细胞内钙有关。总之，这项科学研究开发的方法学不仅是了解肝病病理生理学的有用工具，而且对于肝病的治疗和预防也是有用的工具。

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.