Lin R C, Zhou F C, Fillenwarth M J, Lumeng L
Department of Medicine, Indiana University School of Medicine, Indianapolis.
Hepatology. 1993 Oct;18(4):864-9. doi: 10.1002/hep.1840180418.
Acetaldehyde, a highly reactive intermediate of alcohol metabolism, has been shown to form adducts with liver proteins in rats fed alcohol for long periods. In this report, the zonal distribution of liver protein-acetaldehyde adducts that formed in vivo was studied by means of histoimmunostaining. Rats were pair-fed alcohol-containing and alcohol-free AIN'76 liquid diets for 2 or 11 wk before they were killed and subjected to whole body perfusion with paraformaldehyde. Each liver was cut into 60-microns-thick slices. Slices were first treated with 10% hydrogen peroxide to eliminate endogenous peroxidase activity. They were then incubated sequentially with rabbit antihemocyanin-acetaldehyde adduct, goat antirabbit serum IgG and rabbit peroxidase-antiperoxidase complex. The liver slices were stained with diaminobenzidine and counterstained with methylgreen. In the livers of rats fed alcohol for 2 wk, peroxidase activity was evident in the perivenous zone but not the periportal zone. No staining was obtained when the primary antibody had been preabsorbed with immobilized hemocyanin-acetaldehyde adduct or if the liver slices were incubated with the unimmunized rabbit IgG. Slight staining of the perivenous zone was seen in the livers of control rats, presumably because of minimal protein-acetaldehyde adduct formation emanating from endogenous acetaldehyde. When rats were fed alcohol for longer periods (e.g., 11 wk), protein-acetaldehyde adducts were still seen predominantly in the perivenous zone, but the distribution pattern was more diffuse than that observed in the livers of rats fed alcohol for only 2 wk. More liver cells produced protein-acetaldehyde adducts when rats were fed the alcohol-containing diet supplemented with cyanamide.(ABSTRACT TRUNCATED AT 250 WORDS)
乙醛是酒精代谢的一种高反应性中间体，已证明在长期喂食酒精的大鼠中，它会与肝脏蛋白质形成加合物。在本报告中，通过组织免疫染色研究了体内形成的肝脏蛋白质 - 乙醛加合物的区域分布。大鼠在被处死并用多聚甲醛进行全身灌注之前，成对喂食含酒精和不含酒精的AIN'76液体饮食2周或11周。将每个肝脏切成60微米厚的切片。切片首先用10%过氧化氢处理以消除内源性过氧化物酶活性。然后依次与兔抗血蓝蛋白 - 乙醛加合物、山羊抗兔血清IgG和兔过氧化物酶 - 抗过氧化物酶复合物孵育。肝脏切片用二氨基联苯胺染色，并用甲基绿复染。在喂食酒精2周的大鼠肝脏中，过氧化物酶活性在肝静脉周围区域明显，但在门静脉周围区域未观察到。当一抗用固定化血蓝蛋白 - 乙醛加合物预吸收或肝脏切片与未免疫的兔IgG孵育时，未获得染色。在对照大鼠的肝脏中可见肝静脉周围区域有轻微染色，推测是由于内源性乙醛产生的少量蛋白质 - 乙醛加合物形成。当大鼠长期（例如11周）喂食酒精时，蛋白质 - 乙醛加合物仍主要见于肝静脉周围区域，但分布模式比仅喂食酒精2周的大鼠肝脏中观察到的更弥散。当给大鼠喂食补充了氨甲酰的含酒精饮食时，更多的肝细胞产生了蛋白质 - 乙醛加合物。（摘要截断于250字）
