Pardridge W M, Jefferson L S
Am J Physiol. 1975 Apr;228(4):1155-61. doi: 10.1152/ajplegacy.1975.228.4.1155.
The uptake of 14C-labeled amino acids and carbohydrates by liver following rapid injection into the portal vein was measured relative to a simultaneously injected highly diffusible reference, tritium-labeled water, (3HOH). A 0.25-ml bolus of buffered Ringer solution containing 1-2 muCi of the 14C-labeled test substance and 3-6 muCi of 3HOH was administered by rabid portal injection in anesthetized rats. Circulation was terminated after a single passage of the bolus through the hepatic microvasculature and the tissue was immediately macerated, solubilized, and subjected to liquid scintillation counting. Liver uptake indices (LUI) were calculated from the ratio of 14C to 3H in liver tissue relative to the same ratio in the injection mixture. LUI's of five carbohydrates were measured: sucrose (24.3 percent), inulin (27.7 percent), D-mannitol (80.2 percent), D-glucose (96.8 percent) and L-glucose (26.7 percent). The LUI for cholic acid was 127.1 percent. Among 18 amino acids tested, the LUI's were the highest for the acidic ones (L-aspartic acid, 100.0 percent and L-glutamic acid, 86.4 percent) and lowest for the basic ones (L-arginine, 37.4 percent and L-lysine, 31.4 percent). Stereospecificity for glucose and alanine uptake, saturation kinetics for glutamic acid (Km equal to 4.8 mM) and aspartic acid (Km equal to 2.7 mM), and cross-inhibition among uptake of the acidic amino acids were observed. These findings confirmed the applicability of a technique which was originally developed for studies of amino acid uptake in brain to characterization of transport systems in liver.
通过向门静脉快速注射,测量肝脏对14C标记的氨基酸和碳水化合物的摄取,并与同时注射的高扩散性参考物质氚标记水(3HOH)进行比较。在麻醉大鼠中,通过快速门静脉注射给予0.25毫升含有1 - 2微居里14C标记测试物质和3 - 6微居里3HOH的缓冲林格溶液。在团注单次通过肝微血管后终止循环,立即将组织研磨、溶解并进行液体闪烁计数。肝脏摄取指数(LUI)根据肝脏组织中14C与3H的比例相对于注射混合物中相同比例计算得出。测量了五种碳水化合物的LUI:蔗糖(24.3%)、菊粉(27.7%)、D - 甘露醇(80.2%)、D - 葡萄糖(96.8%)和L - 葡萄糖(26.7%)。胆酸的LUI为127.1%。在测试的18种氨基酸中,酸性氨基酸(L - 天冬氨酸,100.0%和L - 谷氨酸,86.4%)的LUI最高,碱性氨基酸(L - 精氨酸,37.4%和L - 赖氨酸,31.4%)的LUI最低。观察到葡萄糖和丙氨酸摄取的立体特异性、谷氨酸(Km等于4.8 mM)和天冬氨酸(Km等于2.7 mM)的饱和动力学以及酸性氨基酸摄取之间的交叉抑制。这些发现证实了一种最初为研究脑内氨基酸摄取而开发的技术适用于肝脏转运系统的表征。