Suleiman M S, Chapman R A
Department of Physiology, School of Veterinary Science, University of Bristol, United Kingdom.
Cardiovasc Res. 1993 Oct;27(10):1810-4. doi: 10.1093/cvr/27.10.1810.
Many amino acids have been shown to be cotransported with Na+ in a variety of tissues but evidence for an Na+ dependent efflux in the heart is restricted to taurine. The aim of this study was to determine the intracellular levels of the principal amino acids, alanine, glutamine, glutamate, and aspartate, when the heart was beating at normal or low temperature, during exposure to strophanthidin, and when the heart was arrested with either Ca2+,Mg(2+)-free Tyrode solution (a condition known to raise intracellular Na+) or with a high K+ Tyrode solution.
Guinea pig hearts, mounted on a Langendorff apparatus, were perfused with a variety of media and the level of amino acids in the ventricles determined using high performance liquid chromatography.
During perfusion with normal Tyrode solution a time dependent fall in tissue glutamine, glutamate, and aspartate, but not alanine, was observed. Exposure of the beating heart to strophanthidin (0.2 mM) or cooling to 20 degrees C, which should induce a similar increase in [Na+]i but have opposite effects on metabolism, had different effects on tissue amino acids. Cooling did not affect the level of amino acids whereas strophanthidin produced an effect consistent with an activation of a glutamate-alanine aminotransferase. Arresting the heart with Ca2+,Mg(2+)-free Tyrode solution provoked a marked fall in all amino acids which was associated with their appearance in the effluent. This loss of the amino acids was antagonised by conditions known to reduce the rise in intracellular Na+ concentration and increased by inhibition of the Na+ pump. Comparison of the beating heart with the heart arrested with high K+ Tyrode solution showed no difference in the levels of glutamate or aspartate, with a small fall in alanine and taurine and raised glutamine.
The data from beating or arrested hearts are consistent with the presence of amino acid/Na+ cotransport systems but the levels of amino acids free in the sarcoplasm depend on an interaction between amino acid transport driven by the changes in the Na+ gradient and the effects of metabolism.
许多氨基酸已被证明在多种组织中与Na+协同转运,但心脏中依赖Na+的流出证据仅限于牛磺酸。本研究的目的是确定在心脏正常或低温跳动时、暴露于毒毛花苷期间以及用无Ca2+、Mg2+的台氏液(一种已知会升高细胞内Na+的情况)或高K+台氏液使心脏停搏时,主要氨基酸丙氨酸、谷氨酰胺、谷氨酸和天冬氨酸的细胞内水平。
将豚鼠心脏安装在Langendorff装置上,用多种培养基灌注,并使用高效液相色谱法测定心室中的氨基酸水平。
在用正常台氏液灌注期间,观察到组织中的谷氨酰胺、谷氨酸和天冬氨酸随时间下降,但丙氨酸没有。将跳动的心脏暴露于毒毛花苷(0.2 mM)或冷却至20℃,这两种情况应会导致细胞内Na+浓度有类似升高,但对代谢有相反影响,对组织氨基酸有不同影响。冷却不影响氨基酸水平,而毒毛花苷产生的效应与谷氨酸 - 丙氨酸转氨酶的激活一致。用无Ca2+、Mg2+的台氏液使心脏停搏会导致所有氨基酸显著下降,这与它们在流出液中的出现有关。已知能降低细胞内Na+浓度升高的条件可拮抗氨基酸的这种损失,而抑制Na+泵则会增加这种损失。将跳动的心脏与用高K+台氏液停搏的心脏进行比较,发现谷氨酸或天冬氨酸水平没有差异,但丙氨酸和牛磺酸略有下降,谷氨酰胺升高。
来自跳动或停搏心脏的数据与氨基酸/Na+协同转运系统的存在一致,但肌浆中游离氨基酸的水平取决于由Na+梯度变化驱动的氨基酸转运与代谢效应之间的相互作用。