Effects of D-galactosamine (GalN) was studied in primary cultures of adult rat hepatocytes. Hepatocytes were isolated by a in situ collagenase perfusion technique and maintained as monolayers in serum free media for 48 hr. To defect the cytotoxicity, effects of GalN on hormonal induction of α-amino isobutyric acid (AIB) transport system and tyrosine aminotransferase (TAT) activity were studied. Effects on uptake of cardiac glycoside, ouabain was also examined. Treatment of GalN inhibited hormonal induction of AIB uptake markedly. At 0.1 mM of GalN, AIB uptake was inhibited significantly when treated for 12 hr. At higher doses (0.25, 0.5 and 1.0 mM) significant inhibition was noticed after 1 hr exposure. Generally the magnitude of the inhibition was dose and treatment time related. Treatment of GalN to the culture also inhibited hormonal induction of TAT activity. The effects was quite similar pattern as in AIB uptake. Meanwhile, uptake of ouabain was not effected by GalN; up to 1 mM of GalN, uptake of ouabain was similar to that of controls. The viability of the hepatocyte was decreased only slightly by the treatment of GalN; more than 87% of the hepatocytes excluded trypan blue when treated 1 mM GalN for 6 hr. GalN-induced depression of AIB uptake and TAT activity was recovered by the addition of uridine to the culture, simultaneously with GalN or after treatment of GalN.
생화학적으로 바이러스성 간염과 유사한 간염을 일으키는 것으로 알려진 갈락토사민의 독성이 1차 배양 간세포에서 연구 되었다.
1.0mM 농도의 갈락토사민을 1, 3, 6, 및 12시간 처리했을때 세포의 생존률은 87% 이상 이었다.
갈락토사민의 농도를 0.1, 0.25, 0.5, 및 1.0mM 로 1, 3, 6, 및 12시간 처리했을때, 글루카콘 및 덱사메다손에 의해 유도된 AIB 의 세포 배로의 유입량은 갈락토사민의 농도나 처리시간에 따라 현저하게 감소 되었다.
같은 농도와 처리시간으로, ouabain 의 세포내 유입은 영향을 받지 않았으나, 덱사메타손에 의해 유도된 TAT 활성은 현저한 감소를 보였다.
갈락토사민에 의한 AIB 유입량의 감소 및 TAT 활성의 감소는 uridine 을 갈락도사민과 동시에 처리하거나 갈락토사민을 6시간 처리한 후에 처리하였을때 정상 수준의 90% 정도의 회복을 보였다.
