Azumi N
Hokkaido Igaku Zasshi. 1982 May;57(3):331-49.
Cytotoxic effect of excessive copper in the liver of the dogs with longstanding obstructive jaundice were investigated. Common bile duct was ligated in adult mongrel dogs for a period of 21 to 93 days. Copper (0.5 mg/kg weight, every other day) was administered intravenously. Copper content and morphologic changes of the liver was compared with those of the common bile duct ligated dogs without copper administration and of the normal control. Liver copper content was quantitated by atomic absorption spectrophotometry and morphologic investigation was carried out ultrastructurally and histochemically (dimethylaminobenzylidine rhodanine stain for copper and orcein stain for copper associated protein). The copper content of the liver was 57 +/- 8.75 microgram/g wet weight (mean +/- S.E.) in the normal control, 80.84 +/- 15.76 in the common bile duct ligated dogs and 463.46 +/- 76.42 in the common bile duct ligated dogs with copper administration. There was a significant increase (p less than 0.01) of the liver copper content in the common bile duct ligated dogs with copper administration but not in the common bile duct ligated dogs without it. Histologically, the liver showed changes of longstanding cholestasis and of early biliary cirrhosis in the dogs over three months after ligation. Ultrastructurally, both groups showed dilatation of bile canaliculi with decreased and swollen microvilli protruding into their lumina, expanded pericanalicular ectoplasm with increased microfibrils and various forms of intracanalicular and intracytoplasmic bile assuming myelin-figure, crystalloid and dense-amorphous appearances. Also present were increased and dilated smooth-surfaced endoplasmic reticulum, mitochondria showing curled cristae with electron dense ground substance and decreased microvillous projections of hepatocyte cell membranes into Disse's space. Only significant morphologic difference between two groups was the presence of copper-protein complex demonstrated by rhodanine and orcein stains as intracytoplasmic coarse granules in the common bile duct ligated dogs with copper administration. These copper-protein complex granules correspond to partially membrane-bound dense bodies seen ultrastructurally, which probably represent autophagic vacuoles or lysosomal residual bodies. Above result suggests that excessive copper accumulated in the liver as lysosomal bodies in longstanding extrahepatic biliary obstruction with copper loading does not produce significant liver cell injury despite eight fold increase of the liver copper content.
研究了长期梗阻性黄疸犬肝脏中过量铜的细胞毒性作用。将成年杂种犬的胆总管结扎21至93天。静脉注射铜（0.5mg/kg体重，隔日一次）。将肝脏的铜含量和形态学变化与未给予铜的胆总管结扎犬及正常对照犬进行比较。通过原子吸收分光光度法定量肝脏铜含量，并通过超微结构和组织化学方法（用二甲基氨基苄基罗丹宁染色检测铜，用orcein染色检测铜相关蛋白）进行形态学研究。正常对照犬肝脏铜含量为57±8.75μg/g湿重（平均值±标准误），胆总管结扎犬为80.84±15.76，给予铜的胆总管结扎犬为463.46±76.42。给予铜的胆总管结扎犬肝脏铜含量显著增加（p＜0.01），而未给予铜的胆总管结扎犬则无此现象。组织学上，结扎后三个月以上的犬肝脏显示长期胆汁淤积和早期胆汁性肝硬化的变化。超微结构上，两组均显示胆小管扩张，微绒毛减少且肿胀，伸入管腔，胆小管周围胞质扩张，微原纤维增加，管内和胞质内胆汁呈现髓鞘样、晶体样和致密无定形外观。还可见滑面内质网增多和扩张，线粒体嵴卷曲，基质电子密度增加，肝细胞细胞膜向狄氏间隙的微绒毛突起减少。两组之间仅有的显著形态学差异是，给予铜的胆总管结扎犬经罗丹宁和orcein染色显示胞质内有粗大颗粒，即铜-蛋白复合物。这些铜-蛋白复合物颗粒对应于超微结构中部分膜结合的致密小体，可能代表自噬泡或溶酶体残余体。上述结果表明，在长期肝外胆管梗阻且有铜负荷的情况下，肝脏中作为溶酶体体积累的过量铜，尽管肝脏铜含量增加了八倍，但并未产生明显的肝细胞损伤。
