血清蛋白对超灌流大鼠子宫中雌激素介导的受体转位的影响。

Effect of serum proteins on estrogen-mediated receptor translocation in the superfused rat uterus.

作者信息

Reilly J, Rafkind I, Longcope C

机构信息

Department of Obstetrics and Gynecology, University of Massachusetts Medical School, Worcester 01605.

出版信息

J Steroid Biochem. 1987 Dec;28(6):717-22. doi: 10.1016/0022-4731(87)90403-1.

DOI:10.1016/0022-4731(87)90403-1

PMID:3695519

Abstract

In order to examine the effects of serum proteins on the biologic activity of estrogens, we superfused uteri from ovariectomized rats with Krebs-Ringer phosphate buffer (KRP), 4% human serum albumin (HSA) in KR or charcoal-stripped human plasma (HP), alone or with estradiol (E2), estrone (E1) or estriol (E3), 5 x 10(-10), 10(-9) and 10(-8) M. Following superfusion, the uteri were homogenized and the cytosol and nuclear receptors were measured by an exchange technique. Since we could detect no significant difference in the percent of receptors in the nucleus when the time of superfusion was varied from 30-120 min, all studies were done using a 30 min superfusion at a flow rate of 0.6 ml/min. In control studies using KRP alone (n = 12) 23.8 +/- 1.8 (mean +/- SEM) of the receptors were present in the nucleus at the end of the 30 min superfusion. Addition of E1, E2 or E3 5 x 10(-10) M resulted in a significant increase compared to controls in the percent of receptors in the nucleus. The percent of nuclear receptors was significantly greater for E2 and E3 (46.5 +/- 3.2% and 43.6 +/- 1.8%) compared to E1 (34.0 +/- 0.9%). Superfusions of uteri with either E2 or E3 at 10(-9) M or 10(-8) M resulted in a significantly greater percent of nuclear receptors compared to equimolar infusions of E1. When uteri were superfused with E1 at 5 x 10(-10), 10(-9) or 10(-8) M or with E3 at 5 x 10(-10) or 10(-9) M in HSA or HP the percent of nuclear receptors was not different compared to the respective infusion of equimolar concentrations of E1 or E3 in KR. However, superfusions of E2 5 x 10(-10), 10(-9) or 10(-8) M in HSA or HP resulted in a significant decrease in the percent nuclear receptors compared to the percent after equimolar superfusions of E2 in KR. Superfusions of E2 in HSA or HP resulted in the same percent of receptors in the nucleus. The percent of receptors in the nucleus increased with increasing concentrations of E2, but at each concentration the percent of receptors was the same with HA as with HP. Using the percent of nuclear receptors as an index of biological activity, E1 has less activity than either E2 or E3. Interaction with serum proteins does not modulate the activities of either E1 or E3, except at the concentration of 10(-8) M for E3.(ABSTRACT TRUNCATED AT 400 WORDS)

摘要

为了研究血清蛋白对雌激素生物活性的影响，我们用 Krebs - Ringer 磷酸盐缓冲液（KRP）、含 4%人血清白蛋白（HSA）的 KR 溶液或经活性炭处理的人血浆（HP）单独或与雌二醇（E2）、雌酮（E1）或雌三醇（E3）（浓度分别为 5×10⁻¹⁰、10⁻⁹和 10⁻⁸ M）对去卵巢大鼠的子宫进行灌流。灌流后，将子宫匀浆，并用交换技术测量胞质和核受体。由于当灌流时间在 30 - 120 分钟之间变化时，我们未检测到核受体百分比有显著差异，所以所有研究均采用 30 分钟灌流，流速为 0.6 ml/min。在仅使用 KRP 的对照研究中（n = 12），30 分钟灌流结束时，23.8±1.8（均值±标准误）的受体存在于细胞核中。添加 5×10⁻¹⁰ M 的 E1、E2 或 E3 导致细胞核中受体百分比与对照组相比显著增加。与 E1（34.0±0.9%）相比，E2 和 E3 的核受体百分比显著更高（分别为 46.5±3.2%和 43.6±1.8%）。用 10⁻⁹ M 或 10⁻⁸ M 的 E2 或 E3 灌流子宫，导致核受体百分比比等摩尔浓度的 E1 灌流显著更高。当子宫用 5×10⁻¹⁰、10⁻⁹或 10⁻⁸ M 的 E1 或 5×10⁻¹⁰或 10⁻⁹ M 的 E3 在 HSA 或 HP 中灌流时，核受体百分比与在 KR 中相应等摩尔浓度的 E1 或 E3 灌流相比无差异。然而，5×10⁻¹⁰、10⁻⁹或 10⁻⁸ M 的 E2 在 HSA 或 HP 中灌流导致核受体百分比与在 KR 中等摩尔浓度的 E2 灌流后的百分比相比显著降低。E2 在 HSA 或 HP 中灌流导致细胞核中受体百分比相同。细胞核中受体百分比随 E2 浓度增加而增加，但在每个浓度下，HSA 和 HP 中的受体百分比相同。以核受体百分比作为生物活性指标，E1 的活性低于 E2 或 E3。与血清蛋白相互作用除了在 E3 浓度为 10⁻⁸ M 时外，不会调节 E1 或 E3 的活性。（摘要截取自 400 字）