Germinario R J, Oliveira M, Manuel S, Taylor M
Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montreal, Quebec, Canada.
Clin Invest Med. 1987 Jul;10(4):295-302.
The kinetics of saturable and nonsaturable sugar transport were studied in normal and Type II diabetic cultured skin fibroblasts under fast or slowly growing conditions. The Km of hexose transport for fast and slow-growing normal fibroblasts was 1.38 +/- 0.3 and 0.88 +/- 0.12 mM, respectively, while those of the diabetic fibroblasts were 1.57 +/- 0.29 and 0.8 +/- 0.19 mM, respectively. The respective transport Vmax for normal and diabetic fast-growing cells was 13.9 +/- 0.8 and 12.95 +/- 2.4 nmoles 2-DG/mg protein/min. For slowly growing cells of both groups, a transport Vmax of 11.5 +/- 2.4 and 11.3 +/- 1.7 nmoles 2-DG/mg protein/min was obtained. No significant differences were observed in the Km or Vmax of hexose transport under these various growth conditions between normal and diabetic cell cultures. Nonsaturable sugar uptake as determined by L-glucose or cytochalasin B inhibited 2-DG uptake was variable, but no significant differences were observed between the normal and Type II diabetic cells. The activation energies for saturable and nonsaturable sugar uptake were not different among the two donor groups. Insulin stimulation of hexose transport was studied in the presence and absence of dexamethasone (5 X 10(-6) M) at varying insulin concentrations. No difference was observed in the amount of insulin necessary to obtain a maximum stimulatory response (approximately 33 nM insulin in both groups). Also, the insulin concentration required to achieve a one-half maximal response was not significantly different in the donor groups (i.e., 3.53 +/- 0.6 nM for normals and 3.98 +/- 1.1 nM for diabetics.(ABSTRACT TRUNCATED AT 250 WORDS)
在快速生长或缓慢生长条件下,对正常和II型糖尿病培养的皮肤成纤维细胞中可饱和和不可饱和糖转运的动力学进行了研究。快速生长和缓慢生长的正常成纤维细胞的己糖转运Km分别为1.38±0.3和0.88±0.12 mM,而糖尿病成纤维细胞的Km分别为1.57±0.29和0.8±0.19 mM。正常和糖尿病快速生长细胞各自的转运Vmax为13.9±0.8和12.95±2.4 nmol 2-DG/mg蛋白/分钟。对于两组缓慢生长的细胞,转运Vmax为11.5±2.4和11.3±1.7 nmol 2-DG/mg蛋白/分钟。在这些不同生长条件下,正常和糖尿病细胞培养物之间的己糖转运Km或Vmax未观察到显著差异。由L-葡萄糖或细胞松弛素B抑制2-DG摄取所测定的不可饱和糖摄取是可变的,但正常和II型糖尿病细胞之间未观察到显著差异。两个供体组之间可饱和和不可饱和糖摄取的活化能没有差异。在不同胰岛素浓度下,在存在和不存在地塞米松(5×10⁻⁶ M)的情况下研究了胰岛素对己糖转运的刺激作用。在获得最大刺激反应所需的胰岛素量方面未观察到差异(两组中胰岛素约为33 nM)。此外,在供体组中达到最大反应一半所需的胰岛素浓度也没有显著差异(即正常人为3.53±0.6 nM,糖尿病人为3.98±1.1 nM)。(摘要截短至250字)