THE KEY INVOLVEMENT OF ESTROGEN RECEPTOR β AND G-PROTEIN-COUPLED RECEPTOR 30 IN THE NEUROPROTECTIVE ACTION OF DAIDZEIN
THE KEY INVOLVEMENT OF ESTROGEN RECEPTOR β AND G-PROTEIN-COUPLED RECEPTOR 30 IN THE NEUROPROTECTIVE ACTION OF DAIDZEIN
复制标题
DOI：
10.1016/j.neuroscience.2013.02.005
复制
发表时间：
2013-05-15
期刊：
影响因子：
3.3
通讯作者：
Wojtowicz, A. K.
中科院分区：
文献类型：
--
作者：
Kajta, M.;Rzemieniec, J.;Wojtowicz, A. K.
植物雌激素受到了相当多的关注，因为它们提供了一系列有益的作用，例如神经保护。为了更好地了解植物雌激素与经典雌激素受体和膜雌激素受体 (ER) 之间的分子和功能联系，我们研究了大豆黄酮对谷氨酸介导的细胞凋亡途径的影响。我们的研究表明，大豆黄酮（0.1-10 μM）可抑制谷氨酸处理引起的促细胞凋亡和神经毒性作用。海马、新皮质和小脑组织以类似的方式对黄豆苷元对谷氨酸激活的 caspase-3 和乳酸脱氢酶 (LDH) 释放的抑制作用做出反应。 Hoechst 33342 和钙黄绿素 AM 染色在细胞水平上支持生化数据。神经元细胞对大豆苷元介导的保护的敏感性在体外发育早期第 7 天的海马培养物中最为突出。选择性雌激素受体 β (ER β) 拮抗剂 4-[2-苯基-5,7-双(三氟甲基)吡唑并[1,5,-a]嘧啶-3-基]苯酚 (PHTPP) 和选择性 G 蛋白偶联受体 30 (GPR30) 拮抗剂 3aS*， 4R*,9bR*)-4-(6-Bromo-1,3-苯并间二氧杂环己烷-5-基)-3a,4,5,9b-3H-环戊[c]喹啉 (G15)，逆转黄豆苷元介导的谷氨酸诱导的膜线粒体电位、caspase-3 活性和 LDH 释放损失的抑制作用。选择性 ER α 拮抗剂甲基哌啶吡唑 (MPP) 不影响大豆黄酮的任何抗凋亡作用。然而，高亲和力雌激素受体拮抗剂7α,17β-[9-[(4,4,5,5,5-五氟戊基)亚磺酰基]壬基]estra-1,3,5(10)-三烯-3,17-二醇(ICI)182,780和选择性GPR30激动剂， (+/-)-1-[(3aR*,4S*,9bS*)-4-(6-bromo-1,3-苯并间二氧杂环己烷-5-基)-3a,4,5,9b-四氢-3H-环戊[c]喹啉-8-基]-乙酮 (G1)，增强了大豆苷元对谷氨酸诱导的膜线粒体电位损失和 LDH 释放的保护作用。在 siRNA ER beta 和 siRNA GPR30 转染的细胞中，大豆苷元不会抑制谷氨酸诱导的作用。暴露于谷氨酸 24 小时不会影响 ER β 和 GPR30 的细胞分布，但导致受体水平增加超过 100%。与大豆苷元共同处理降低了 ER β 水平，但 GPR30 蛋白水平没有显着变化。在这里，我们阐明了低微摩尔浓度的大豆黄酮的神经保护作用，并证明植物雌激素可能通过新型核外 GPR30 和经典的转录作用 ER beta 发挥其作用。这些研究揭示了 ER beta 和 GPR30 细胞内信号通路在介导大豆黄酮抗凋亡作用中的关键作用，并可能为治疗或预防神经变性的新策略提供见解。 (C) 2013 国际广播组织。由爱思唯尔有限公司出版。保留所有权利。
Phytoestrogens have received considerable attention because they provide an array of beneficial effects, such as neuroprotection. To better understand the molecular and functional link between phytoestrogens and classical as well as membrane estrogen receptors (ERs), we investigated the effect of daidzein on the glutamate-mediated apoptotic pathway. Our study demonstrated that daidzein (0.1-10 mu M) inhibited the pro-apoptotic and neurotoxic effects caused by glutamate treatment. Hippocampal, neocortical and cerebellar tissues responded to the inhibitory action of daidzein on glutamate-activated caspase-3 and lactate dehydrogenase (LDH) release in a similar manner. Biochemical data were supported at the cellular level by Hoechst 33342 and calcein AM staining. The sensitivity of neuronal cells to daidzein-mediated protection was most prominent in hippocampal cultures at an early stage of development 7th day in vitro. A selective estrogen receptor beta (ER beta) antagonist, 4-[2-phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5,-a]pyrimidin-3-yl]phenol (PHTPP), and a selective G-protein-coupled receptor 30 (GPR30) antagonist, 3aS*, 4R*,9bR*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H- cyclopenta[c]quinoline (G15), reversed the daidzein-mediated inhibition of glutamate-induced loss of membrane mitochondrial potential, caspase-3 activity, and LDH release. A selective ER alpha antagonist, methyl-piperidino-pyrazole (MPP), did not influence any anti-apoptotic effect of daidzein. However, a high-affinity estrogen receptor antagonist, 7 alpha,17 beta-[9-[(4,4,5,5,5-pentafluoropentyl)sulfinyl]nonyl]estra-1,3,5(10)-triene-3,17-diol (ICI) 182,780, and a selective GPR30 agonist, (+/-)-1-[(3aR*,4S*,9bS*)-4-(6-bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethanone (G1), intensified the protective action of daidzein against glutamate-induced loss of membrane mitochondrial potential and LDH release. In siRNA ER beta- and siRNA GPR30-transfected cells, daidzein did not inhibit the glutamate-induced effects. Twenty-four hour exposure to glutamate did not affect the cellular distribution of ER beta and GPR30, but caused greater than 100% increase in the levels of the receptors. Co-treatment with daidzein decreased the level of ER beta without significant changing of the GPR30 protein level. Here, we elucidated neuroprotective effects of daidzein at low micromolar concentrations and demonstrated that the phytoestrogens may exert their effects through novel extranuclear GPR30 and the classical transcriptionally acting ER beta. These studies uncover key roles of the ER beta and GPR30 intracellular signaling pathways in mediating the anti-apoptotic action of daidzein and may provide insight into new strategies to treat or prevent neural degeneration. (C) 2013 IBRO. Published by Elsevier Ltd. All rights reserved.