Effects of tibolone and its metabolites on Angiopoietin-1, Tie-2 and tumor necrosis factor-at mRNA in Ishikawa cells implication for tibolone's effects on the endometrium
Effects of tibolone and its metabolites on Angiopoietin-1, Tie-2 and tumor necrosis factor-at mRNA in Ishikawa cells implication for tibolone's effects on the endometrium
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DOI：
10.1016/j.maturitas.2007.03.002
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发表时间：
2007-08-20
期刊：
影响因子：
4.9
通讯作者：
Archer, David R.
中科院分区：
文献类型：
--
作者：
Mirkin, Sebastian;Archer, David R.
目的：确定17 β-雌二醇、醋酸甲羟孕酮、替勃龙和替勃龙代谢产物对体外石川细胞中血管生成素-1、Tie-2和肿瘤坏死因子-a的影响。我们假设，个别激素对血管生成因子或炎性细胞因子的差异效应可能与绝经后妇女使用激素therapy.Design子宫内膜出血有关：石川细胞培养至80%汇合，在体外。在无血清培养基中孵育24小时后，将1.0、0.1和0.01 μ M的17 β-雌二醇、醋酸甲羟孕酮、替勃龙、3 α-羟基替勃龙、3 β-羟基替勃龙和δ（4）-替勃龙加入到石川细胞中。然后将细胞加类固醇再孵育24小时。从对照和处理的石川细胞中提取总RNA。逆转录后，在掺入33 p-dCTP的聚合酶链反应中扩增血管生成素-1、Tie-2、肿瘤坏死因子-α和β-肌动蛋白cDNA。Angiopoietin-1，Tie-2和肿瘤坏死因子-α mRNA的相对丰度测定闪烁spectroscopic.Results：17 β-雌二醇和醋酸甲羟孕酮增加Angiopoietin-1 mRNA显着高于对照组，替勃龙和替勃龙羟基代谢产物。在所有测试浓度下的Δ（4）-替勃龙均不增加血管生成素-1。与对照组相比，在任何浓度下测试的类固醇都没有改变Tie-2 mRNA表达。1.0和0.1 μ M的17-β-雌二醇使肿瘤坏死因子-α mRNA显著高于对照组。醋酸甲羟孕酮仅在1.0 μ M时增加肿瘤坏死因子-α mRNA高于对照水平。Tibolone、3 α-羟基Tibolone、3 β-羟基Tibolone和Delta（4）-Tibolone在每种浓度下对肿瘤坏死因子-α mRNA丰度没有影响。结论：Delta（4）-Tibolone不刺激Angiopoietin-1，而其他类固醇具有比对照更大的差异效应。激素对Tie-2 mRNA表达无明显影响。17 β-雌二醇和最高浓度的醋酸甲羟孕酮可增加肿瘤坏死因子-α。我们解释这些结果支持我们的假设，即个别类固醇激素对血管生成因子或炎症细胞因子的不同作用可能与使用激素治疗的绝经后妇女子宫内膜出血的临床发生有关。(c)2007爱思唯尔爱尔兰有限公司保留所有权利。
Objective: To determine the effect of 17 beta-estradiol, medroxyprogesterone acetate, tibolone and tibolone metabolites on Angiopoietin-1, Tie-2, and tumor necrosis factor-a in Ishikawa cells, in vitro. We hypothesized that differential effects on angiogenic factors or inflammatory cytokines by individual hormones may be related to the endometrial bleeding in postmenopausal women using hormone therapy.Design: Ishikawa cells were cultured to 80% confluence, in vitro. After 24h incubation in serum-free media, 1.0, 0.1 and 0.01 mu M of 17 beta-estradiol, medroxyprogesterone acetate, tibolone, 3 alpha-hydroxytibolone, 3 beta-hydroxytibolone, and Delta(4)-tibolone were added to the Ishikawa cells. The cells plus steroids were then incubated for a further 24 h. Total RNA was extracted from control and treated Ishikawa cells. After reverse transcription, Angiopoietin-1, Tie-2, tumor necrosis factor-alpha, and beta-actin cDNAs were amplified in a polymerase chain reaction spiked with 33p-dCTP. Relative abundance of Angiopoietin-1, Tie-2, and tumor necrosis factor-alpha mRNA was measured by scintillation spectroscopy.Results: 17 beta-Estradiol and medroxyprogesterone acetate increased Angiopoietin-1 mRNA significantly higher than control, tibolone and tibolone hydroxy metabolites. Delta(4)-Tibolone at all concentrations tested did not increase Angiopoietin-1. None of the steroids tested at any concentration altered Tie-2 mRNA expression compared to control. 17 P-Estradiol at 1.0 and 0.1 mu M increased tumor necrosis factor-alpha mRNA significantly higher than control. Medroxyprogesterone acetate only at 1.0 mu M increased tumor necrosis factor-alpha mRNA above control levels. Tibolone, 3 alpha-hydroxytibolone, 3 beta-hydroxytibolone, and Delta(4)-tibolone at every concentration had no effect on tumor necrosis factor-alpha mRNA abundance.Conclusions: Delta(4)-Tibolone did not stimulate Angiopoietin-1, while the other steroids had differential effects greater than control. None of the steroids changed the expression of Tie-2 mRNA. Tumor necrosis factor-alpha was increased by 17 beta-estradiol and by the highest concentration of medroxyprogesterone acetate. We interpret these results as supportive of our hypothesis that differential effects on angiogenic factors or inflammatory cytokines by individual steroids may be related to the clinical occurrence of endometrial bleeding in postmenopausal women using hormone therapy. (c) 2007 Elsevier Ireland Ltd. All rights reserved.