Juel C, Wibrand F
Zoophysiological Laboratory B, August Krogh Institute, Copenhagen, Denmark.
Acta Physiol Scand. 1989 Sep;137(1):33-9. doi: 10.1111/j.1748-1716.1989.tb08718.x.
Lactate transport across the sarcolemma of isolated mouse muscles was studied with a 14C tracer technique. The cellular tracer uptake could be inhibited by unlabelled L-lactate (and pyruvate) and to a lesser extent by D-lactase. The stereospecific fraction had a Km of 3.5 mM, and made up 50% of the total transport. The tracer uptake was unaffected by 0.05 mM DIDS and 0.2 mM amiloride, but was inhibited by cinnamate (Ki = 8 mM) and PCMBS (Ki = 0.8 mM). With high concentrations of the latter inhibitor compounds or with high concentrations of unlabelled L-lactate, the tracer uptake was inhibited 80%, which indicates that the main part of the transport involves facilitated diffusion. The remaining fraction (20%) was non-saturable, reduced at high pH, and could not be inhibited; it is probably mediated by diffusion of undissociated lactic acid. Lactate transport was pH-dependent, which is consistent with a lactate-H+ symport. The maximal transport capacity, as calculated from the pH changes measured with pH-sensitive micro-electrodes while the lactate gradient was 30 mM, was 11.8 mmol kg-1 min-1 (pH 6.2).
采用¹⁴C示踪技术研究了乳酸在分离的小鼠肌肉肌膜上的转运。未标记的L-乳酸(和丙酮酸)可抑制细胞对示踪剂的摄取,D-乳糖的抑制作用较小。立体特异性部分的Km为3.5 mM,占总转运量的50%。示踪剂摄取不受0.05 mM DIDS和0.2 mM氨氯吡咪的影响,但受肉桂酸盐(Ki = 8 mM)和对氯汞苯甲酸(Ki = 0.8 mM)抑制。使用高浓度的后一种抑制剂化合物或高浓度的未标记L-乳酸时,示踪剂摄取被抑制80%,这表明转运的主要部分涉及易化扩散。其余部分(20%)不可饱和,在高pH值下减少,且不能被抑制;它可能是由未解离乳酸的扩散介导的。乳酸转运依赖于pH值,这与乳酸-H⁺同向转运一致。当乳酸梯度为30 mM时,根据用pH敏感微电极测量的pH变化计算出的最大转运能力为11.8 mmol kg⁻¹ min⁻¹(pH 6.2)。