Mori M, Yamaguchi J, Oyamada M, Enomoto K, Kaneko A
Department of Pathology, Sapporo Medical College, Japan.
Cell Struct Funct. 1990 Oct;15(5):263-71. doi: 10.1247/csf.15.263.
Horseradish peroxidase (HRP, 10 mg/100 g body weight) was intravenously injected into rats in order to investigate the nature of the compartments involved in the transcellular transport of the protein through hepatocytes into bile. Double cytochemistry for HRP and the marker enzymes for cytoplasmic organelles was used. HRP was shown to be taken up by hepatocytes via vesicles at the sinusoidal surface, some of which were positive for 5'-nucleotidase activity. HRP was then found in the smooth-surfaced vesicles and tubules which were negative in 5'-nucleotidase, glucose 6-phosphatase, thiamine pyrophosphatase and acid phosphatase activity, suggesting that the tubular structures are neither the endoplasmic reticulum, the Golgi apparatus nor lysosomes. Biochemical studies revealed that the lead procedures used for the double cytochemistry did not inhibit the peroxidatic activity of HRP, and conversely that HRP did not interfere with the marker enzyme activity. Such cytochemical observations seemed to be supported by the observation that administration of monensin (3.5 mg/100 g) and chloroquine (5 mg/100 g), which markedly altered the structure of the Golgi apparatus and lysosomes, respectively, slightly altered the biliary excretion of HRP but not to a significant extent.
为了研究蛋白质通过肝细胞跨细胞转运至胆汁过程中所涉及的区室的性质，将辣根过氧化物酶（HRP，10毫克/100克体重）静脉注射到大鼠体内。采用了HRP与细胞质细胞器标记酶的双重细胞化学方法。结果显示，HRP通过肝血窦表面的小泡被肝细胞摄取，其中一些小泡对5'-核苷酸酶活性呈阳性。随后在5'-核苷酸酶、葡萄糖6-磷酸酶、硫胺素焦磷酸酶和酸性磷酸酶活性呈阴性的光滑表面小泡和小管中发现了HRP，这表明这些管状结构既不是内质网、高尔基体也不是溶酶体。生化研究表明，用于双重细胞化学的前处理步骤不会抑制HRP的过氧化物酶活性，反之，HRP也不会干扰标记酶的活性。分别显著改变高尔基体和溶酶体结构的莫能菌素（3.5毫克/100克）和氯喹（5毫克/100克）给药后，HRP的胆汁排泄略有改变但程度不显著，这一观察结果似乎支持了上述细胞化学观察。
