Lopaschuk G D, Witters L A, Itoi T, Barr R, Barr A
Department of Pediatrics, University of Alberta, Edmonton, Canada.
J Biol Chem. 1994 Oct 14;269(41):25871-8.
Fatty acid oxidation rapidly increases in the rabbit heart following birth. By inhibiting carnitine palmitoyltransferase 1 (CPT1), malonyl-CoA is a potent regulator of fatty acid oxidation in the heart. We therefore addressed the hypothesis that a decrease in acetyl-CoA carboxylase (ACC) activity and/or malonyl-CoA inhibition of CPT1 could account for the increase in the ability of the heart to oxidize fatty acids following birth. ACC activity and expression, malonyl-CoA levels, and mitochondrial CPT1 activity were measured in hearts from 1-day, 7-day, and 6-week-old rabbits. CPT1 activity and sensitivity to malonyl-CoA inhibition did not differ between 1-day, 7-day, or 6-week hearts (the IC50 for malonyl-CoA was 32.0 +/- 1.5, 36.0 +/- 0.3, and 36.3 nM, respectively). Western blot analysis with streptavidin showed that all hearts expressed similar amounts of both a 265-kDa (ACC-265) and 280-kDa isoform (ACC-280) of ACC. A progressive and significant decrease in malonyl-CoA levels was seen in 1-day, 7-day, and 6-week hearts (47 +/- 2, 40 +/- 2, and 26 +/- 2 nmol/g dry weight, respectively), paralleling a decline in ACC activity. We hypothesized that these developmental changes could be due to changes in hormonal regulation of cardiac ACC in the postnatal period. In isolated hearts from 1-day-old rabbits, the fatty acid oxidation rate was 9.01 +/- 1.10 nmol.g dry weight-1.min-1. Glucagon (1 ng/ml) did not alter this rate (11.03 +/- 1.42 nmol.g dry weight-1.min-1), but insulin (100 microunits/ml) resulted in a significant decrease in rate (4.81 +/- 0.82 nmol.g dry weight-1.min-1). ACC activity was markedly elevated in 1-day-old hearts perfused with insulin compared to control hearts or glucagon perfused hearts (0.415 +/- 0.052, 0.095 +/- 0.018, and 0.133 +/- 0.013 nmol of malonyl-CoA produced.g dry weight-1.min-1, respectively). Malonyl-CoA levels were also markedly elevated in 1-day hearts perfused with insulin (123.0 +/- 8.3, 2.0 +/- 0.4, and 1.8 +/- 0.6 nmol/g dry weight in insulin, control, and glucagon hearts, respectively). In 7-day-old rabbit hearts, the basal fatty acid oxidation rate had increased to 24.5 +/- 4.8 nmol.mg-1.min-1. In contrast to the 1-day-old hearts, insulin had no significant effect on fatty acid oxidation, although glucagon resulted in a significant increase in rates (38.9 +/- 12.2 and 80.7 +/- 9.1 nmol.g dry weight-1.min-1, respectively).(ABSTRACT TRUNCATED AT 400 WORDS)
出生后,家兔心脏中的脂肪酸氧化迅速增加。丙二酰辅酶A通过抑制肉碱棕榈酰转移酶1(CPT1),成为心脏中脂肪酸氧化的有效调节剂。因此,我们探讨了以下假说:乙酰辅酶A羧化酶(ACC)活性降低和/或丙二酰辅酶A对CPT1的抑制作用,可能是出生后心脏脂肪酸氧化能力增加的原因。我们测定了1日龄、7日龄和6周龄家兔心脏中的ACC活性和表达、丙二酰辅酶A水平以及线粒体CPT1活性。1日龄、7日龄或6周龄心脏之间的CPT1活性以及对丙二酰辅酶A抑制的敏感性并无差异(丙二酰辅酶A的半数抑制浓度分别为32.0±1.5、36.0±0.3和36.3 nM)。用链霉亲和素进行的蛋白质印迹分析表明,所有心脏中ACC的265 kDa(ACC-265)和280 kDa亚型(ACC-280)的表达量相似。在1日龄、7日龄和6周龄心脏中,丙二酰辅酶A水平逐渐且显著降低(分别为47±2、40±2和[26±2 nmol/g干重]),这与ACC活性的下降平行。我们推测,这些发育变化可能是由于出生后心脏ACC激素调节的改变。在1日龄家兔的离体心脏中,脂肪酸氧化速率为9.01±1.10 nmol·g干重⁻¹·min⁻¹。胰高血糖素(1 ng/ml)并未改变该速率(11.03±1.42 nmol·g干重⁻¹·min⁻¹),但胰岛素(100微单位/ml)导致速率显著降低(4.81±0.82 nmol·g干重⁻¹·min⁻¹)。与对照心脏或灌注胰高血糖素的心脏相比,灌注胰岛素的1日龄心脏中ACC活性显著升高(分别为0.415±0.052、0.095±0.018和0.133±0.013 nmol丙二酰辅酶A生成·g干重⁻¹·min⁻¹)。灌注胰岛素的1日龄心脏中丙二酰辅酶A水平也显著升高(胰岛素、对照和胰高血糖素心脏中的丙二酰辅酶A水平分别为123.0±8.3、2.0±0.4和[1.8±0.6 nmol/g干重])。在7日龄家兔心脏中,基础脂肪酸氧化速率已增加至24.5±4.8 nmol·mg⁻¹·min⁻¹。与1日龄心脏不同,胰岛素对脂肪酸氧化无显著影响,尽管胰高血糖素导致速率显著增加(分别为38.9±12.2和80.7±9.1 nmol·g干重⁻¹·min⁻¹)。(摘要截选至400字)