Moseley R H, Murphy S M
Department of Internal Medicine, Veterans Administration Medical Center, University of Michigan School of Medicine, Ann Arbor 48105.
Am J Physiol. 1989 Mar;256(3 Pt 1):G458-65. doi: 10.1152/ajpgi.1989.256.3.G458.
Ethanol has been reported to inhibit hepatocellular processes such as gluconeogenesis and protein synthesis that depend, in part, on amino acid uptake. Since previous studies in cultured hepatocytes indicate that ethanol may have a direct and selective inhibitory effect on amino acid transport, the effects of ethanol on amino acid uptake into basolateral (sinusoidal) rat liver plasma membrane (blLPM) vesicles were examined. Uptake of [3H]alanine, [3H]leucine, and [35S]cysteine was measured by a rapid Millipore filtration technique in the presence of inwardly directed Na+ and K+ gradients and under tetramethylammonium (TMA+)- and Na+-equilibrated conditions. Ethanol preincubation produced a concentration-dependent inhibition of Na+-dependent alanine and cysteine uptake; no effect was observed on either Na+-independent alanine and cysteine uptake or Na+-independent leucine transport. Ethanol had no effect on L-alanine transport under Na+-equilibrated conditions; however, initial rates of 22Na flux were enhanced in the presence of ethanol. On the basis of differences in 2-(methylamino)isobutyrate and L-cysteine sensitivity, ethanol inhibition of Na+-dependent alanine transport in blLPM vesicles largely but not exclusively corresponded to the hormone-responsive system A for amino acid transport described in isolated hepatocytes. Kinetic analysis showed that ethanol treatment resulted in an alteration in the apparent maximum velocity of reaction (Vmax) of Na+-dependent alanine transport without affecting the apparent Km for alanine. The inhibitory effects of ethanol on the time course of Na+-dependent alanine uptake were reversible.(ABSTRACT TRUNCATED AT 250 WORDS)
据报道,乙醇会抑制肝细胞过程,如糖异生和蛋白质合成,这些过程部分依赖于氨基酸摄取。由于先前在培养肝细胞中的研究表明,乙醇可能对氨基酸转运有直接和选择性的抑制作用,因此研究了乙醇对大鼠肝基底外侧(窦状隙)质膜(blLPM)囊泡摄取氨基酸的影响。通过快速密理博过滤技术,在存在内向的Na⁺和K⁺梯度以及在四甲基铵(TMA⁺)和Na⁺平衡条件下,测量[³H]丙氨酸、[³H]亮氨酸和[³⁵S]半胱氨酸的摄取。乙醇预孵育对依赖Na⁺的丙氨酸和半胱氨酸摄取产生浓度依赖性抑制;对不依赖Na⁺的丙氨酸和半胱氨酸摄取或不依赖Na⁺的亮氨酸转运均未观察到影响。在Na⁺平衡条件下,乙醇对L-丙氨酸转运无影响;然而,在乙醇存在下,²²Na通量的初始速率增加。基于2-(甲氨基)异丁酸和L-半胱氨酸敏感性的差异,乙醇对blLPM囊泡中依赖Na⁺的丙氨酸转运的抑制在很大程度上但并非完全对应于分离肝细胞中描述的氨基酸转运的激素反应性系统A。动力学分析表明,乙醇处理导致依赖Na⁺的丙氨酸转运反应的表观最大速度(Vmax)发生改变,而不影响丙氨酸的表观Km。乙醇对依赖Na⁺的丙氨酸摄取时间进程的抑制作用是可逆的。(摘要截短于250字)