Rapid effects of androgens in macrophages
Rapid effects of androgens in macrophages
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DOI：
10.1016/j.steroids.2004.05.007
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发表时间：
2004-08-01
期刊：
影响因子：
2.7
通讯作者：
Wunderlich, F
中科院分区：
文献类型：
--
作者：
Benten, WPM;Guo, Z;Wunderlich, F
我们研究了小鼠巨噬细胞IC-21和RAW 264.7中睾酮膜受体（mAR）的存在及其在非基因组途径、基因表达和细胞功能中的作用。这两种细胞系都缺乏细胞内雄激素受体（iARs），对睾丸激素的反应是[Ca2+]的快速上升(i)。这些[Ca2+](i)的升高既不能被iAR-也不能被iER阻滞剂抑制，而是通过mAR介导。药理学方法表明，mAR属于通过百日破毒素（PTX）敏感的g蛋白与磷脂酶C偶联的膜受体。通过共聚焦激光扫描显微镜（CLSM）和流式细胞术可以定位为睾酮- bsa - fitc的特异性表面结合位点，并且具有激动剂隔离性。为了研究睾酮诱导的[Ca2+]升高对基因表达的可能作用，将c-fos启动子报告基因构建物转染到RAW 264.7巨噬细胞中。[Ca2+](i)的增加，诱导睾酮不能直接显著激活c-fos启动子。此外，单独睾酮刺激后，未观察到ERK1/2、JNK/SAPK和p38的显著激活。然而，睾酮诱导的[Ca2+](i)升高确实对脂多糖（LPS）诱导的遗传信号传导背景下的基因表达有特定影响：睾酮特异性下调脂多糖诱导的c-fos启动子、p38 MAPK和NO生成的激活。在胎牛血清（FCS）诱导的基因向信号传导中，情况正好相反，即睾酮增加了c-fos启动子和ERK1/2的激活。我们的研究表明，在巨噬细胞中，睾酮诱导的非基因组Ca2+信号与LPS和FCS诱导的基因性信号之间存在交叉。(C) 2004爱思唯尔公司版权所有。
We investigated the existence of membrane receptors for testosterone (mAR) in mouse macrophages of the cell lines IC-21 and RAW 264.7 as well as their roles in nongenomic pathways, gene expression and cell functioning. Both cell lines lack intracellular androgen receptors (iARs) and respond to testosterone with rapid rises in [Ca2+](i). These rises in [Ca2+](i) can neither be inhibited by iAR- nor by iER blockers, but are rather mediated through mAR. Pharmacological approaches suggest that the mAR belongs to the class of membrane receptors which are coupled to phospholipase C via pertussis toxin (PTX) sensitive G-proteins. The mAR can be localized as specific surface binding sites for testosterone-BSA-FITC by confocal laser scanning microscopy (CLSM)and flow cytometry, and are characterized by their agonist-sequestrability. In order to examine a possible role of the testosterone-induced rise in [Ca2+], on gene expression, a c-fos promoter reporter gene construct was transfected into RAW 264.7 macrophages. The increase in [Ca2+](i), induced testosterone cannot significantly activate the c-fos promoter directly. Also, no significant activation of ERK1/2, JNK/SAPK and p38 can be observed following testosterone-stimulation alone. However, testosterone-induced rises in [Ca2+](i) do have specific effects on gene expression in context with lipopolysaccharide (LPS)-induced genotropic signaling: testosterone specifically down-regulates LPS-induced activation of c-fos promoter, p38 MAPK and NO production. In fetal calf serum (FCS)-induced genotropic signaling, the situation is reversed, i.e. testosterone augments the activation of c-fos promoter and ERK1/2. Our studies demonstrate a cross-talk between the testosterone-induced nongenomic Ca2+ signaling and the genotropic signaling induced by LPS and FCS in macrophages. (C) 2004 Elsevier Inc. All rights reserved.