Sakaguchi K, Fukase M, Kobayashi I, Fujita T
Department of Medicine, Kobe University School of Medicine, Japan.
J Bone Miner Res. 1986 Jun;1(3):259-65. doi: 10.1002/jbmr.5650010304.
The effect of parathyroid hormone (PTH) on the time course of glucose-6-phosphate dehydrogenase (G6PD) activity in the distal convoluted tubule of a vitamin D-depleted guinea pig was determined using quantitative cytochemistry. G6PD activity decreased to the stable basal level 5 hrs after the initiation of the kidney segment maintenance cultures. The exposure of the tissues to 1 pg/ml of bovine PTH-(1-84) induced a cyclic change of G6PD activity, whereas neither carboxyl-terminal PTH nor other hormones tested showed such activity. After a 16-min exposure to bovine PTH-(1-84), the peak height of each cycle began to decrease until it disappeared at 34 min. The second exposure to this hormone at 46 min reinduced a similar cyclic change with a similar peak, indicating full viability of the cells. When bovine PTH-(1-84) was incubated with an excess amount of anti-bovine PTH antibody, the PTH-induced G6PD activity was completely abolished. Throughout a 14-min exposure to either human PTH-(1-84), human PTH-(1-34) or bovine PTH-(1-84), similar cyclic changes were observed with the constant peak height regardless of the dose (10(-16)-10(-12) M), although the cycle length shortened progressively as the dose was increased. They were equipotent on a molar basis between the concentrations of 10(-16) and 10(-13) M at 6 min of hormone exposure. The present data demonstrate that the cytochemical bioassay of PTH in a vitamin D-depleted animal is based on a dose-dependent difference in the time course of G6PD activity.
利用定量细胞化学方法,测定了甲状旁腺激素(PTH)对维生素D缺乏的豚鼠远曲小管中葡萄糖-6-磷酸脱氢酶(G6PD)活性随时间变化的影响。肾段维持培养开始5小时后,G6PD活性降至稳定的基础水平。将组织暴露于1 pg/ml的牛PTH-(1-84)会诱导G6PD活性发生周期性变化,而羧基末端PTH或其他测试激素均未表现出这种活性。在暴露于牛PTH-(1-84)16分钟后,每个周期的峰值高度开始下降,直至在34分钟时消失。在46分钟时再次暴露于该激素会再次诱导出类似的具有相似峰值的周期性变化,表明细胞具有完全的活力。当牛PTH-(1-84)与过量的抗牛PTH抗体孵育时,PTH诱导的G6PD活性完全消失。在长达14分钟的时间内,无论是暴露于人PTH-(1-84)、人PTH-(1-34)还是牛PTH-(1-84),均观察到类似的周期性变化,且峰值高度恒定,与剂量(10^-16 - 10^-12 M)无关,尽管随着剂量增加,周期长度逐渐缩短。在激素暴露6分钟时,它们在10^-16至10^-13 M的浓度范围内摩尔等效。目前的数据表明,在维生素D缺乏的动物中,PTH的细胞化学生物测定是基于G6PD活性随时间变化的剂量依赖性差异。