Burger H J, Gebhardt R, Mayer C, Mecke D
Physiologisch-Chemisches Institut, Universität Tübingen, Federal Republic of Germany.
Hepatology. 1989 Jan;9(1):22-8. doi: 10.1002/hep.1840090105.
Periportal and perivenous hepatocytes were isolated from rat liver by digitonin/collagenase perfusion for investigating the acinar heterogeneity of amino acid transport activities related to glutamine and ammonia metabolism. Immunocytochemical staining of the respective subpopulations for glutamine synthetase demonstrated that periportal subpopulations were essentially free of glutamine synthetase-positive cells, whereas perivenous subpopulations showed a 2- to 3-fold enrichment of glutamine synthetase-positive hepatocytes. The high perivenous/periportal ratio of 59 found for glutamine synthetase activity as well as the perivenous/periportal ratios of other marker enzymes further indicated the good separation of periportal and perivenous cells. alpha-Aminoisobutyric acid, histidine and glutamate were used to determine the distribution pattern of amino acid transport systems A, N and G-, as well as of the sodium-independent uptake of these compounds 1 hr after isolation and after maximal hormonal stimulation during primary culture. The strong heterogeneity of the sodium-independent transport of histidine, characterized by higher perivenous transport rates [perivenous/periportal ratio: 1.5 (1 hr) to 3.5 (48 hr)], suggests a significant role of facilitated diffusion, presumably in glutamine export. Conversely, the strong heterogeneity of the sodium-dependent glutamate transport (System G-) characterized by higher uptake rates in nonstimulated [perivenous/periportal ratio: 6.6 (1 hr)] and in hormonally treated perivenous hepatocytes (perivenous/periportal ratio: 2.2) reflects its possible significance with respect to the substrate availability for glutamine synthesis. The observed heterogeneities provide a basis for understanding how substrate fluxes related to glutamine metabolism might be established and regulated.(ABSTRACT TRUNCATED AT 250 WORDS)
通过洋地黄皂苷/胶原酶灌注从大鼠肝脏中分离出汇管区和中央静脉周围的肝细胞,以研究与谷氨酰胺和氨代谢相关的氨基酸转运活性的腺泡异质性。对各亚群进行谷氨酰胺合成酶的免疫细胞化学染色显示,汇管区亚群基本没有谷氨酰胺合成酶阳性细胞,而中央静脉周围亚群的谷氨酰胺合成酶阳性肝细胞富集了2至3倍。谷氨酰胺合成酶活性的中央静脉周围/汇管区高比率59以及其他标记酶的中央静脉周围/汇管区比率进一步表明汇管区和中央静脉周围细胞得到了良好分离。使用α-氨基异丁酸、组氨酸和谷氨酸来确定氨基酸转运系统A、N和G-的分布模式,以及这些化合物在分离后1小时和原代培养期间最大激素刺激后的非钠依赖性摄取情况。组氨酸非钠依赖性转运的强烈异质性,其特征是中央静脉周围转运速率较高[中央静脉周围/汇管区比率:1.5(1小时)至3.5(48小时)],表明促进扩散可能在谷氨酰胺输出中起重要作用。相反,钠依赖性谷氨酸转运(系统G-)的强烈异质性,其特征是在未刺激的[中央静脉周围/汇管区比率:6.6(1小时)]和经激素处理的中央静脉周围肝细胞(中央静脉周围/汇管区比率:2.2)中摄取率较高,反映了其在谷氨酰胺合成底物可用性方面的可能意义。观察到的异质性为理解与谷氨酰胺代谢相关的底物通量如何建立和调节提供了基础。(摘要截短于250字)