Bernus I, Mitchell A M, Manley S W, Mortimer R H
Division of Chemical Pathology, Queensland Health Pathology Service, Royal Brisbane Hospital Campus, Australia.
Placenta. 1999 Mar-Apr;20(2-3):161-5. doi: 10.1053/plac.1998.0358.
This study investigated uptake of triiodothyronine sulphate (T3S) and interactions between uptake of T3S and triiodothyronine (T3) using the human choriocarcinoma cell line (JAr) as a model of placental transport. Cells were incubated at 37 degrees C with 30 pM 125I-T3 for 2 min with unlabelled T3 (0-30 microM) or T3S (0-1 mM). Addition of an excess unlabelled T3 (30 microM) or T3S (1 mM) reduced the initial rate of 125I-T3 uptake by 69.3+/-3.6 per cent (P<0.0001) and 52.9+/-7.8 per cent (P<0.0001), respectively. The calculated Michaelis constant (Km) for T3 uptake was 0.378+/-0.133 microM (n = 3) with a corresponding maximum velocity (Vmax) of 15.4+/-6.9 pmol/min/mg protein. Uptake of 125I-T3 was inhibited in a dose-dependent way by the addition of unlabelled T3S (0-1 mM). The calculated inhibition constant (Ki) for the inhibition of 125I-T3 uptake by T3S was 121.8+/-35.2 microM (n = 6). Saturable uptake of 125I-T3S by JAr cells was negligible. The T3S preparation incubated with the cells contained about 0.1 per cent T3, sufficient to explain the apparent inhibition of 125I-T3 uptake by unlabelled T3S. These results suggest that, in contrast to T3 uptake in these cells, JAr cells do not have a saturable uptake mechanism for T3S, and that T3S does not interact with the T3 transporter in these cells.
本研究以人绒毛膜癌细胞系(JAr)作为胎盘转运模型,研究了硫酸三碘甲状腺原氨酸(T3S)的摄取以及T3S与三碘甲状腺原氨酸(T3)摄取之间的相互作用。将细胞在37℃下与30 pM 125I-T3孵育2分钟,同时加入未标记的T3(0 - 30 microM)或T3S(0 - 1 mM)。加入过量的未标记T3(30 microM)或T3S(1 mM)分别使125I-T3的初始摄取速率降低了69.3±3.6%(P<0.0001)和52.9±7.8%(P<0.0001)。计算得出T3摄取的米氏常数(Km)为0.378±0.133 microM(n = 3),相应的最大速度(Vmax)为15.4±6.9 pmol/分钟/毫克蛋白。加入未标记的T3S(0 - 1 mM)以剂量依赖性方式抑制125I-T3的摄取。T3S抑制125I-T3摄取的计算抑制常数(Ki)为121.8±35.2 microM(n = 6)。JAr细胞对125I-T3S的可饱和摄取可忽略不计。与细胞一起孵育的T3S制剂含有约0.1%的T3,足以解释未标记的T3S对125I-T3摄取的明显抑制作用。这些结果表明,与这些细胞中T3的摄取不同,JAr细胞没有T3S的可饱和摄取机制,并且T3S在这些细胞中不与T3转运体相互作用。