Segal J, Ingbar S H
J Clin Invest. 1979 Mar;63(3):507-15. doi: 10.1172/JCI109329.
Studies were conducted to ascertain the in vitro effect of 3,5,3'-triiodothyronine (T(3)) on the accumulation of the glucose analogue, 2-deoxyglucose (2-DG), by thymocytes freshly isolated from weanling rats. At a concentration of 1 muM, T(3) stimulated the 15-min uptake of (3)H-2-DG after cells had been exposed to T(3) for only 30 min. Significant stimulation of 2-DG accumulation was produced by 1 nM T(3), with increasing stimulation at doses ranging up to 10 muM. T(3) did not alter the fraction of accumulated 2-DG that was phosphorylated, and kinetic studies indicated that its effect was associated with a significant increase in the apparent V(max) of 2-DG accumulation, but not the apparent K(m). T(3) also enhanced the accumulation by thymocytes of the nonmetabolized glucose analogue, 3-O-methylglucose (3-O-MG), an effect that was evidently the result of an increase in 3-O-MG transport into the cell, because it was seen in cells incubated with (3)H-3-O-MG for only 30 s. The proportionate increase in 2-DG accumulation produced by T(3) was not altered by preincubating cells with concentrations of puromycin or cycloheximide sufficient to reduce [(3)H]-leucine incorporation by 95%, and T(3) over a period of >2 h had no effect on [(3)H]leucine incorporation itself. These results indicate that T(3) stimulates the uptake of sugars in rat thymocytes in vitro by an effect on their inward transport. The promptness of the effect and its failure to be inhibited during profound inhibition of protein synthesis further indicate that this effect of T(3) is not mediated through a nuclear-dependent mechanism. Rather, the properties of this response, and of the increases in amino acid and 2-DG accumulation produced by T(3) in other tissue preparations, strongly suggest that these effects of T(3) are mediated at the level of cell membrane.
开展了多项研究,以确定3,5,3'-三碘甲状腺原氨酸(T3)对从断奶大鼠新鲜分离的胸腺细胞积累葡萄糖类似物2-脱氧葡萄糖(2-DG)的体外作用。在浓度为1μM时,细胞仅暴露于T3 30分钟后,T3就刺激了(3)H-2-DG的15分钟摄取。1 nM T3产生了对2-DG积累的显著刺激,在高达10μM的剂量范围内刺激作用增强。T3没有改变积累的2-DG中被磷酸化的部分,动力学研究表明其作用与2-DG积累的表观V(max)显著增加有关,但与表观K(m)无关。T3还增强了胸腺细胞对非代谢葡萄糖类似物3-O-甲基葡萄糖(3-O-MG)的积累,这种作用显然是3-O-MG转运进入细胞增加的结果,因为在与(3)H-3-O-MG孵育仅30秒的细胞中就观察到了这种作用。用足以使[(3)H]-亮氨酸掺入减少95%的嘌呤霉素或环己酰亚胺浓度预孵育细胞,不会改变T3产生的2-DG积累的相应增加,并且在超过2小时的时间段内T3对[(3)H]亮氨酸掺入本身没有影响。这些结果表明,T3通过影响大鼠胸腺细胞的内向转运来刺激糖的摄取。该作用的迅速性以及在蛋白质合成受到深度抑制期间未被抑制,进一步表明T3的这种作用不是通过核依赖机制介导的。相反,这种反应的特性以及T3在其他组织制剂中产生的氨基酸和2-DG积累增加的特性,强烈表明T3的这些作用是在细胞膜水平介导的。