Changes odf hapatic sinusoidal cell in fibrotic process of chronic liver disease-Three dimendional examination by confocal laser scanning microscopy-

部分用抗α-平滑肌肌动蛋白（αSMA）抗体对正常人肝和肝硬化患者肝组织（50 μm厚）进行免疫组化染色，并用激光共聚焦扫描显微镜观察，发现α SMA阳性星状细胞在正常人肝小叶内分布较均匀，胞质突起呈三维延伸，包围肝窦。α SMA-反应性在再生结节之间变化。与正常肝组织相比，再生结节中的星状细胞胞质突起的α SMA免疫反应性降低，并被胞质突起包围，胞质突起呈二维延伸。此外，α SMA阳性的星状细胞密集地排列在纤维间隔中，并且不通过胞质突起包围血窦腔。用抗胶质细胞酸性蛋白（GFAP）抗体对大鼠正常肝组织进行免疫组化染色，观察ET-1（1或400 nmol/l）灌注对肝星状细胞的影响。GFAP阳性星状细胞与正常人肝内α SMA阳性星状细胞的整体观相同，胞质突起呈三维延伸。此外，他们往往有较长的细胞质突起之间的区域门静脉和中央周围地区。与仅用缓冲液灌注的星形细胞相比，ET-1灌注导致GFAP阳性的星形细胞胞体肿胀，中间丝浓缩，但未观察到星形细胞胞质突起明显缩短。从这些结果，它表明，在正常肝脏中的星状细胞不参与的正弦流。

Sections (50-μm thick) obtained from normal and cirrhotic human livers were immunostained with anti-α smooth muscle actin (αSMA) antibody and were examined by confocal laser scanning microscopy, αSMA-positive stellate cells were distributed relatively evenly in the hepatic lobules of normal human livers and surrounded the sinusoids by the cytoplasmic processes showing three-dimensional (3-D) extension, On the other hand, αSMA-reactivity varied between regenerative nodules in cirrhotic livers. Stellate cells in the regenerative nodules showed reduction of αSMA-immunoreactivity of the cytoplasmic processes compared with that in normal livers and surrounded the sinusoids by cytoplasmic processes with two-dimensional extensions. In addition, αSMA-positive stellate cells were densely packed in fibrous septa and did not surround the lumens of the sinusoid by the cytoplasmic processes. These results showed that stellate cells have different characteristics compared with those in the normal liver.In rats, the sections from normal livers with and without ET-1 perfusion (1 or 400 nmol/l) were immunostained with anti-glial fibrillary acidic protein (GFAP) antibody. GFAP-positive stellate cells showed the entire views same as αSMA-positive stellate cells in the normal human liver and had the cytoplasmic processes with 3-D extensions. Furthermore, they tended to have longer cytoplasmic processes in the area between periportal and pericentral areas. Perfusion of ET-1 resulted in swelling of cell bodies with condensation of the intermediate filaments in GFAP-positive stellate cells compared with those perfused with buffer only However no apparent shortening of the cytoplasmic processes of the stellate cells was observed. From these results, it is suggested that stellate cells in the normal liver are not involved in the sinusoidal flow.