Department of Pathobiochemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka, 569-1094, Japan.
Department of Pathobiochemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka, 569-1094, Japan.
Toxicology. 2021 Feb 28;450:152686. doi: 10.1016/j.tox.2021.152686. Epub 2021 Jan 21.
6-Hydroxydopamine (6-OHDA) is a neurotoxin that destroy dopaminergic neurons and widely used to establish animal models of Parkinson's disease. Prostaglandins (PGs) are involved in various cellular processes, including the damage and repair of neuronal cells. However, the function of PGF in neuronal cells remains unclear. In this study, we investigated the effects of PGF against 6-OHDA-mediated toxicity in human neuroblastoma SH-SY5Y cells and elucidated its underlying molecular mechanism. When the cells were treated with 6-OHDA (50 μM) for 6 h, the expression levels of PGF synthetic enzymes; cyclooxygenase-2 and aldo-keto reductase 1C3 as PGF synthase were enhanced in an incubation-time-dependent manner. In addition, the production of PGF was increased in 6-OHDA-treated cells. Fluprostenol, a PGF receptor (FP) agonist (500 nM), suppressed 6-OHDA-induced cell death by decreasing the production of reactive oxygen species (ROS) and increasing the expression of the anti-oxidant genes. These fluprostenol-mediated effects were inhibited by co-treatment with AL8810, an FP receptor antagonist (1 μM) or transfection with FP siRNA (20 nM). Moreover, 6-OHDA-induced phosphorylation of extracellular signal-regulated kinase (ERK), a member of the mitogen-activated protein kinase family, was inhibited by co-incubation with AL8810. Furthermore, fluprostenol itself enhanced ERK phosphorylation and further elevated the 6-OHDA-induced phosphorylation of ERK. In addition, 6-OHDA induced nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2), activating anti-oxidant gene expression, was repressed by co-culturing with AL8810. These results indicate that PGF suppressed 6-OHDA-induced neuronal cell death by enhancing anti-oxidant gene expression via the FP receptor-ERK-Nrf2 signaling. Thus, FP receptor is a potential target for inhibition of ROS-mediated neuronal cell death.
6-羟多巴胺（6-OHDA）是一种神经毒素，可破坏多巴胺能神经元，广泛用于建立帕金森病的动物模型。前列腺素（PGs）参与各种细胞过程，包括神经元细胞的损伤和修复。然而，PGF 在神经元细胞中的功能尚不清楚。在这项研究中，我们研究了 PGF 对人神经母细胞瘤 SH-SY5Y 细胞中 6-OHDA 介导的毒性的影响，并阐明了其潜在的分子机制。当细胞用 6-OHDA（50 μM）孵育 6 小时时，PGF 合成酶的表达水平；环加氧酶-2 和醛酮还原酶 1C3 作为 PGF 合酶呈孵育时间依赖性增强。此外，在 6-OHDA 处理的细胞中 PGF 的产生增加。氟前列醇，PGF 受体（FP）激动剂（500 nM），通过降低活性氧（ROS）的产生和增加抗氧化基因的表达来抑制 6-OHDA 诱导的细胞死亡。这些氟前列醇介导的作用被 FP 受体拮抗剂（AL8810，1 μM）或 FP siRNA（20 nM）共转染抑制。此外，6-OHDA 诱导的细胞外信号调节激酶（ERK）的磷酸化，ERK 是丝裂原活化蛋白激酶家族的成员，被 AL8810 共孵育抑制。此外，氟前列醇本身增强 ERK 磷酸化，并进一步升高 6-OHDA 诱导的 ERK 磷酸化。此外，6-OHDA 诱导核转录因子红系 2 相关因子 2（Nrf2）的核易位，激活抗氧化基因表达，被 AL8810 共培养抑制。这些结果表明，PGF 通过 FP 受体-ERK-Nrf2 信号通路增强抗氧化基因表达，抑制 6-OHDA 诱导的神经元细胞死亡。因此，FP 受体是抑制 ROS 介导的神经元细胞死亡的潜在靶点。
