Graduate Program in Biotechnology, University of Vale do Taquari (Univates), Lajeado, RS 95914-014, Brazil.
Department of Pharmaceutical and Medicinal Chemistry, Institute of Pharmacy, Eberhard Karls Universität Tübingen, D-72076 Tübingen, Germany.
Int J Mol Sci. 2021 Apr 2;22(7):3701. doi: 10.3390/ijms22073701.
Current treatments for neurodegenerative diseases (ND) are symptomatic and do not affect disease progression. Slowing this progression remains a crucial unmet need for patients and their families. c-Jun N-terminal kinase 3 (JNK3) are related to several ND hallmarks including apoptosis, oxidative stress, excitotoxicity, mitochondrial dysfunction, and neuroinflammation. JNK inhibitors can play an important role in addressing neuroprotection. This research aims to evaluate the neuroprotective, anti-inflammatory, and antioxidant effects of a synthetic compound (FMU200) with known JNK3 inhibitory activity in SH-SY5Y and RAW264.7 cell lines. SH-SY5Y cells were pretreated with FMU200 and cell damage was induced by 6-hydroxydopamine (6-OHDA) or hydrogen peroxide (HO). Cell viability and neuroprotective effect were assessed with an MTT assay. Flow cytometric analysis was performed to evaluate cell apoptosis. The HO-induced reactive oxygen species (ROS) generation and mitochondrial membrane potential (ΔΨm) were evaluated by DCFDA and JC-1 assays, respectively. The anti-inflammatory effect was determined in LPS-induced RAW264.7 cells by ELISA assay. In undifferentiated SH-SY5Y cells, FMU200 decreased neurotoxicity induced by 6-OHDA in approximately 20%. In RA-differentiated cells, FMU200 diminished cell death in approximately 40% and 90% after 24 and 48 h treatment, respectively. FMU200 reduced both early and late apoptotic cells, decreased ROS levels, restored mitochondrial membrane potential, and downregulated JNK phosphorylation after HO exposure. In LPS-stimulated RAW264.7 cells, FMU200 reduced TNF-α levels after a 3 h treatment. FMU200 protects neuroblastoma SH-SY5Y cells against 6-OHDA- and HO-induced apoptosis, which may result from suppressing the JNK pathways. Our findings show that FMU200 can be a useful candidate for the treatment of neurodegenerative disorders.
目前用于治疗神经退行性疾病（ND）的方法是对症治疗，无法延缓疾病的进展。减缓疾病进展仍然是患者及其家属的一个重要未满足的需求。c-Jun N 末端激酶 3（JNK3）与几种 ND 的特征有关，包括细胞凋亡、氧化应激、兴奋性毒性、线粒体功能障碍和神经炎症。JNK 抑制剂在神经保护中可以发挥重要作用。本研究旨在评估具有已知 JNK3 抑制活性的合成化合物（FMU200）在 SH-SY5Y 和 RAW264.7 细胞系中的神经保护、抗炎和抗氧化作用。SH-SY5Y 细胞用 FMU200 预处理，然后用 6-羟多巴胺（6-OHDA）或过氧化氢（HO）诱导细胞损伤。通过 MTT 测定评估细胞活力和神经保护作用。通过流式细胞术分析评估细胞凋亡。通过 DCFDA 和 JC-1 测定分别评估 HO 诱导的活性氧（ROS）生成和线粒体膜电位（ΔΨm）。通过 ELISA 测定评估 LPS 诱导的 RAW264.7 细胞中的抗炎作用。在未分化的 SH-SY5Y 细胞中，FMU200 降低了 6-OHDA 诱导的神经毒性，约为 20%。在 RA 分化的细胞中，FMU200 在 24 和 48 小时处理后分别使细胞死亡减少约 40%和 90%。FMU200 减少了早晚期凋亡细胞，降低了 ROS 水平，恢复了线粒体膜电位，并下调了 HO 暴露后的 JNK 磷酸化。在 LPS 刺激的 RAW264.7 细胞中，FMU200 在 3 小时处理后降低了 TNF-α 水平。FMU200 可保护神经母细胞瘤 SH-SY5Y 细胞免受 6-OHDA 和 HO 诱导的细胞凋亡，这可能是由于抑制了 JNK 途径。我们的研究结果表明，FMU200 可能是治疗神经退行性疾病的有用候选药物。
