Koho Ninna M, Raekallio Marja, Kuusela Erja, Vuolle Jaana, Pösö A Reeta
Department of Basic Veterinary Sciences, University of Helsinki, POB 66, FIN-00014 Helsinki, Finland.
Am J Vet Res. 2008 Aug;69(8):1091-6. doi: 10.2460/ajvr.69.8.1091.
To detect monocarboxylate transporters (MCTs) in canine RBC membranes and to determine the distribution of lactate between plasma and RBCs.
Blood samples obtained from 6 purpose-bred Beagles.
Monocarboxylate transporter isoforms 1, 2, 4, 6, 7, and 8 and CD147 were evaluated in canine RBCs by use of western blot analysis. Lactate influx into RBCs was measured as incorporation of radioactive lactate.
2 MCT isoforms, MCT1 and MCT7, were detected in canine RBC membranes on western blot analysis, whereas anti-MCT2, anti-MCT4, anti-MCT6, and anti-MCT8 antibodies resulted in no signal. No correlation was found between the amount of MCT1 or MCT7 and lactate transport activity, but the ancillary protein CD147 that is needed for the activity of MCT1 had a positive linear correlation with the rate of lactate influx. The apparent Michael is constant for the lactate influx in canine RBCs was 8.8 +/- 0.9mM. Results of in vitro incubation studies revealed that at lactate concentrations of 5 to 15mM, equilibrium of lactate was rapidly obtained between plasma and RBCs.
These results indicated that at least half of the lactate transport in canine RBCs occurs via MCT1, whereas MCT7 may be responsible for the rest, although an additional transporter was not ruled out. For practical purposes, the rapid equilibration of lactate between plasma and RBCs indicated that blood lactate concentrations may be estimated from plasma lactate concentrations.
检测犬红细胞膜中的单羧酸转运体(MCTs),并确定血浆与红细胞之间乳酸的分布情况。
从6只专门培育的比格犬获取血样。
通过蛋白质印迹分析评估犬红细胞中的单羧酸转运体亚型1、2、4、6、7和8以及CD147。以放射性乳酸的掺入量来测定乳酸流入红细胞的情况。
蛋白质印迹分析在犬红细胞膜中检测到2种MCT亚型,即MCT1和MCT7,而抗MCT2、抗MCT4、抗MCT6和抗MCT8抗体未产生信号。未发现MCT1或MCT7的量与乳酸转运活性之间存在相关性,但MCT1活性所需的辅助蛋白CD147与乳酸流入速率呈正线性相关。犬红细胞中乳酸流入的表观米氏常数为8.8±0.9mM。体外孵育研究结果显示,在乳酸浓度为5至15mM时,血浆与红细胞之间迅速达到乳酸平衡。
这些结果表明,犬红细胞中至少一半的乳酸转运通过MCT1进行,而MCT7可能负责其余部分,尽管不排除存在其他转运体。实际应用中,血浆与红细胞之间乳酸的快速平衡表明可根据血浆乳酸浓度估算血乳酸浓度。