Rapid effects of 17β-estradiol on cell signaling and function of Mytilus hemocytes
Rapid effects of 17β-estradiol on cell signaling and function of Mytilus hemocytes
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DOI：
10.1016/j.ygcen.2003.12.003
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发表时间：
2004-03-01
影响因子：
2.7
通讯作者：
Gallo, G
中科院分区：
文献类型：
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作者：
Canesi, L;Ciacci, C;Gallo, G
雌激素影响几个非生殖组织的功能，特别是免疫系统。在哺乳动物免疫细胞中，17β-雌二醇(E-2)具有剂量和细胞型的特异性效应，对E2的反应似乎是由快速的非基因组机制介导的；这些机制可以在膜或胞浆位置启动，并可以导致直接的局部效应，如离子通量的改变，以及继而激活不同的激酶级联反应的基因转录调节，包括丝裂原激活蛋白激酶(MAPKs)。在这项工作中，我们研究了雌激素对双壳类贝类血细胞、免疫细胞的短期效应及其可能的细胞信号转导机制。结果表明，E-2(25 NM)可引起血细胞胞浆[Ca~(2+)]迅速显著升高，较低浓度(5 NM)对[Ca~(2+)]的影响较小，但不显著。在加入E-2后的5-15分钟内，两种浓度的E-2都会影响酪氨酸激酶介导的信号转导MAPK和STAT样蛋白的磷酸化状态。25 nM E-2的作用更强，时程更清晰：尤其是E-2可引起p-ERK2 MAPK的一过性增加和p-p38 MAPK的持续增加。此外，STAT3和STAT5在E-2作用下均被酪氨酸磷酸化。E-2(5 NM)在加入后10-30分钟内引起形态和功能的变化(如水解酶的胞外释放、溶酶体膜的不稳定和杀菌活性的刺激)。两种浓度的E-2诱导的溶酶体膜失稳可被p38 MAPK抑制剂SB203580预先孵育血细胞所消除，而被PD98059和Wortmannin(分别为ERK MAPK和PI3-K的抑制剂)显著减轻，这表明激酶级联的快速激活参与了E-2对贻贝血细胞的影响。抗雌激素他莫昔芬阻止或强烈降低了E-2的大部分作用，但不是全部。异源抗ERα-抗ERβ抗体的Western blotting显示，在血细胞蛋白提取液中存在免疫反应性的ERα和ERβ样蛋白。总体而言，我们的数据支持这样一种假设，即17β-雌二醇的快速效应和作用机制是极其保守的，它们可能在无脊椎动物的内分泌-免疫相互作用中发挥关键作用。(C)2003 Elsevier Inc.保留所有权利。
Estrogens affect the functioning of several non-reproductive tissues, the immune system in particular. In mammalian immunocytes, 17beta-estradiol (E-2) has both dose- and cell-type specific effects and the responses to E2 seem to be mediated by rapid, nongenomic mechanisms; these may be initiated at either membrane or cytosolic locations, and can result in both direct local effects, such as modification of ion fluxes, and regulation of gene transcription secondary to activation of different kinase cascades, including mitogen activated protein kinases (MAPKs). In this work, the short-term effects of E2 and the possible mechanisms of estrogen-mediated cell signaling were investigated in the hemocytes, the immune cells of the bivalve mollusc, the mussel Mytilus galloprovincialis Lam. The results show that E-2 (25 nM) caused a rapid and significant increase in hemocyte cytosolic [Ca2+]; lower concentrations (5 nM) showed a smaller, not significant effect. Both E-2 concentrations affected the phosphorylation state of the components of tyrosine kinase-mediated signal transduction MAPK- and STAT- (signal transducers and activators of transcription) like proteins within 5-15 min from E-2 addition. A greater effect and clearer time course were observed with 25 nM E-2: in particular, E-2 induced a transient increase in p-ERK2 MAPK and a persistent increase in p-p38 MAPK. Moreover, both STAT3 and STAT5 were tyrosine phosphorylated in response to E-2. E-2 (5 nM) induced both morphological (as evaluated by SEM) and functional changes (such as extracellular release of hydrolytic enzymes, lysosomal membrane destabilisation, and stimulation of the bactericidal activity) within 10-30 min from addition. Lysosomal membrane destabilisation induced by both E-2 concentrations was abolished by hemocyte preincubation with the p38 MAPK inhibitor SB203580, and significantly reduced by PD98059 and Wortmannin (inhibitors of ERK MAPK and PI3-K, respectively), this suggesting that rapid activation of kinase cascades is involved in mediating the effects of E-2 in mussel hemocytes. The antiestrogen Tamoxifen prevented or strongly reduced most, but not all, the effects of E-2. Western blotting with heterologous anti-ERalpha-anti-ERbeta-antibodies revealed the presence of immunoreactive ERalpha- and ERbeta-like proteins in hemocyte protein extracts. Overall, our data support the hypothesis that the rapid effects and mechanisms of action of 17beta-estradiol are extremely conserved and that they may play a crucial role in endocrine-immune interactions in invertebrates. (C) 2003 Elsevier Inc. All rights reserved.