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作为糖皮质激素靶组织的胎盘:(3H)-地塞米松结合的交换测定及类固醇对受体含量的影响。

The placenta as a glucocorticoid target tissue: exchange assay of (3H)-dexamethasone binding and effect of steroids on receptor content.

作者信息

Heller C, Coirini H, De Nicola A F

出版信息

Horm Metab Res. 1983 Jan;15(1):41-5. doi: 10.1055/s-2007-1018622.

Abstract

Binding of (3H)-dexamethasone was studied in high speed supernatant (HSS) of basal zone trophoblast and labyrinthine zone of rat placenta using an exchange assay. The system showed the following characteristics: (1) maximum binding was attained after 120 min of incubation; (2) molybdate was required in the medium to measure binding sites at 20 degrees C and equilibrium conditions; (3) exchange was near 90-100% between ligand and corticosterone added during a preincubation; (4) low capacity (190-250 fmoles/mg protein), high affinity (Kd 10(-8) M) binders were determined by saturation analysis; (5) competition with other steroids in vitro revealed that in basal zone trophoblast, only dexamethasone displaced the ligand, whereas in labyrinthine zone corticosterone, progesterone and testosterone competed to a smaller degree than dexamethasone. We have also studied the effect of in vivo treatment of pregnant rats for four days with dexamethasone, corticosterone, estradiol, progesterone or testosterone, on (3H)-dexamethasone binding. Of all tested compounds, only dexamethasone treatment (which was suspended 24 h before the experiment to prevent receptor occupancy by exogenous hormone) significantly decreased binding of the tritiated hormone in HSS of labyrinthine zone and basal zone trophoblast. These results suggest that dexamethasone regulates its own receptor in placenta. The physiological aspect of this phenomenon requires elucidation of the role of glucocorticoids on placental function in the rat.

摘要

采用交换分析法研究了(3H)-地塞米松在大鼠胎盘基底层滋养层和迷路层高速上清液(HSS)中的结合情况。该系统具有以下特点:(1)孵育120分钟后达到最大结合;(2)在20℃和平衡条件下,培养基中需要钼酸盐来测量结合位点;(3)预孵育期间添加的配体与皮质酮之间的交换接近90-100%;(4)通过饱和分析确定低容量(190-250飞摩尔/毫克蛋白质)、高亲和力(Kd 10(-8)M)的结合剂;(5)体外与其他类固醇的竞争显示,在基底层滋养层中,只有地塞米松能置换配体,而在迷路层中,皮质酮、孕酮和睾酮的竞争程度比地塞米松小。我们还研究了对怀孕大鼠进行为期四天的地塞米松、皮质酮、雌二醇、孕酮或睾酮体内治疗对(3H)-地塞米松结合的影响。在所有测试化合物中,只有地塞米松治疗(在实验前24小时停用,以防止外源性激素占据受体)显著降低了迷路层和基底层滋养层HSS中氚化激素的结合。这些结果表明地塞米松在胎盘中调节其自身受体。这一现象的生理学方面需要阐明糖皮质激素在大鼠胎盘功能中的作用。

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