Burgaya F, Peinado J, Vilaró S, Llobera M, Ramírez I
Departament de Bioquímica i Fisiologia, Facultat de Biologia, Universitat de Barcelona, Spain.
Biochem J. 1989 Apr 1;259(1):159-66. doi: 10.1042/bj2590159.
The lipoprotein lipase activity in the liver of neonatal (1 day old) rats was about 3 times that in the liver of adult rats. Perfusion of the neonatal liver with collagenase decreased the tissue-associated activity by 77%. When neonatal-rat liver cells were dispersed, hepatocyte-enriched (fraction I) and haemopoietic-cell-enriched (fraction II) populations were obtained. The lipoprotein lipase activity in fraction I was 7 times that in fraction II. On the basis of those activities and the proportion of both cell types in either fraction, it was estimated that hepatocytes contained most, if not all, the lipoprotein lipase activity detected in collagenase-perfused neonatal-rat livers. From those calculations it was also concluded that haemopoietic cells did not contain lipoprotein lipase activity. When the hepatocyte-enriched cell population was incubated at 25 degrees C for up to 3 h, a slow but progressive release of enzyme activity to the incubation medium was found. However, the total activity (cells + medium) did not significantly change through the incubation period. Cycloheximide produced a time-dependent decrease in the cell-associated activity. Heparin increased the amount of lipoprotein lipase activity released to the medium. Because the cell-associated activity was unchanged, heparin also produced a time-dependent increase in the total activity. In those cells incubated with heparin, cycloheximide did not affect the initial release of lipoprotein lipase activity to the medium, but blocked further release. The cell-associated activity was also decreased by the presence of cycloheximide in those cells. It is concluded that neonatal-rat hepatocytes synthesize active lipoprotein lipase.
新生(1日龄)大鼠肝脏中的脂蛋白脂肪酶活性约为成年大鼠肝脏的3倍。用胶原酶灌注新生大鼠肝脏可使组织相关活性降低77%。当分散新生大鼠肝细胞时,可获得富含肝细胞的部分(I部分)和富含造血细胞的部分(II部分)。I部分中的脂蛋白脂肪酶活性是II部分的7倍。根据这些活性以及任一分离部分中两种细胞类型的比例,估计肝细胞包含了在胶原酶灌注的新生大鼠肝脏中检测到的大部分(如果不是全部)脂蛋白脂肪酶活性。从这些计算中还得出结论,造血细胞不含有脂蛋白脂肪酶活性。当将富含肝细胞的细胞群体在25℃下孵育长达3小时时,发现酶活性缓慢但逐渐释放到孵育培养基中。然而,在整个孵育期间,总活性(细胞 + 培养基)没有显著变化。放线菌酮使细胞相关活性呈时间依赖性降低。肝素增加了释放到培养基中的脂蛋白脂肪酶活性量。由于细胞相关活性未改变,肝素也使总活性呈时间依赖性增加。在与肝素一起孵育的那些细胞中,放线菌酮不影响脂蛋白脂肪酶活性向培养基中的初始释放,但阻止了进一步释放。在那些细胞中,放线菌酮的存在也降低了细胞相关活性。结论是新生大鼠肝细胞合成活性脂蛋白脂肪酶。