脂肪组织中代谢物浓度的测量以及胰岛素、四氧嘧啶糖尿病和肾上腺素的影响。

Measurement of concentrations of metabolites in adipose tissue and effects of insulin, alloxan-diabetes and adrenaline.

作者信息

Denton R M, Yorke R E, Randle P J

出版信息

Biochem J. 1966 Aug;100(2):407-19. doi: 10.1042/bj1000407.

DOI:10.1042/bj1000407

PMID:5968539

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1265150/

Abstract

Methods are described for the extraction and assay of ATP, ADP, AMP, glucose 6-phosphate, l-glycerol 3-phosphate and citrate in rat epididymal adipose tissue incubated in vitro for 1hr. At this time of incubation rates of glucose uptake and outputs of glycerol, free fatty acids, lactate and pyruvate were shown to be constant. 2. In fat pads incubated in medium containing glucose (3mg./ml.) and albumin (20mg./ml.) the concentrations (in mmumoles/g. wet wt.) were: ATP, 70; ADP, 36; AMP, 9.0; glucose 6-phosphate, 3.0; l-glycerol 3-phosphate, 3.3; citrate, 8.1. 3. The volume of intracellular water calculated from ([(3)H]water space-[(14)C]sorbitol space), ([(14)C]urea space-inulin space) and (weight loss on drying-[(14)C]sorbitol space) was 1.4ml./100g. wet wt. of tissue. The intracellular volume was not changed by insulin, alloxan-diabetes or adrenaline. 4. When compared in terms of mumoles/ml. of intracellular water the concentration of ATP in adipose tissue was less than in heart and diaphragm muscles. The concentrations of ADP and AMP were greater both in absolute terms and relative to ATP. Insulin, alloxan-diabetes and adrenaline had no significant effects on the concentrations of the adenine nucleotides in adipose tissue. 5. The concentration of glucose 6-phosphate was increased by insulin and lowered by alloxan-diabetes and adrenaline. The concentration of l-glycerol 3-phosphate was increased by insulin, unchanged by alloxan-diabetes and lowered by adrenaline. The concentration of citrate was increased by adrenaline and alloxan-diabetes and unchanged by insulin. 6. The effect of glucose concentration in the medium on rates of glucose uptake in adipose tissue from normal rats and alloxan-diabetic rats was investigated. The K(u) of glucose uptake was 29-44mg./100ml. and the V(max.) was 0.77mg./g. wet wt. of tissue/hr. Insulin increased the V(max.) and alloxan-diabetes diminished it, but neither agent significantly altered the K(u). 7. The significance of these results in relation to control of metabolism of adipose tissue is discussed.

摘要

本文描述了体外培养1小时的大鼠附睾脂肪组织中ATP、ADP、AMP、6-磷酸葡萄糖、3-磷酸甘油和柠檬酸的提取及测定方法。在该培养时间，葡萄糖摄取率以及甘油、游离脂肪酸、乳酸和丙酮酸的输出率均保持恒定。2. 在含有葡萄糖（3mg/ml）和白蛋白（20mg/ml）的培养基中培养的脂肪垫中，（以微摩尔/克湿重计）各物质浓度分别为：ATP，70；ADP，36；AMP，9.0；6-磷酸葡萄糖，3.0；3-磷酸甘油，3.3；柠檬酸，8.1。3. 根据（[³H]水空间 - [¹⁴C]山梨醇空间）、（[¹⁴C]尿素空间 - 菊粉空间）以及（干燥失重 - [¹⁴C]山梨醇空间）计算得出的细胞内水体积为1.4ml/100g组织湿重。胰岛素、四氧嘧啶糖尿病或肾上腺素对细胞内体积无影响。4. 就微摩尔/毫升细胞内水而言，脂肪组织中ATP的浓度低于心脏和膈肌肌肉。ADP和AMP的浓度无论绝对值还是相对于ATP的相对值均更高。胰岛素、四氧嘧啶糖尿病和肾上腺素对脂肪组织中腺嘌呤核苷酸的浓度无显著影响。5. 胰岛素可使6-磷酸葡萄糖浓度升高，四氧嘧啶糖尿病和肾上腺素则使其降低。胰岛素可使3-磷酸甘油浓度升高，四氧嘧啶糖尿病对其无影响，肾上腺素则使其降低。肾上腺素和四氧嘧啶糖尿病可使柠檬酸浓度升高，胰岛素对其无影响。6. 研究了培养基中葡萄糖浓度对正常大鼠和四氧嘧啶糖尿病大鼠脂肪组织葡萄糖摄取率的影响。葡萄糖摄取的K(u)为29 - 44mg/100ml，V(max)为0.77mg/g组织湿重/小时。胰岛素可提高V(max)，四氧嘧啶糖尿病则使其降低，但二者均未显著改变K(u)。7. 讨论了这些结果与脂肪组织代谢控制的相关性。