Department of Neurology, Daejeon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Clinical Research Institute, Daejeon St. Mary's Hospital, Daejeon, Republic of Korea.
Biochem Biophys Res Commun. 2022 Sep 17;621:59-66. doi: 10.1016/j.bbrc.2022.06.099. Epub 2022 Jul 1.
Heat shock proteins (HSPs) play an essential role as molecular chaperones to prevent abnormal protein aggregation and misfolding. Moreover, they protect dopamine neurons from oxidative stress, inflammation, and apoptosis, all well-known pathomechanisms of Parkinson's disease (PD). Melatonin is a potent antioxidant that has the beneficial ability to prevent neurodegenerative diseases like PD. We aimed to explore the protective properties of melatonin in an in vitro PD model, focusing on its underlying mechanism using HSPs. A 1-methyl-4-phenylpyridimium (MPP+)-induced toxin model was established with retinoic acid (RA)-differentiated SH-SY5Y cells. Cell viability and apoptosis were measured using MTT and DAPI. Intracellular reactive oxygen species (ROS) levels were measured by the cell-permeant fluorescent probe DCFH-DA. The level of malondialdehyde and the activities of superoxide dismutase and glutathione peroxidase were assessed using ELISA kits. Apoptotic markers of Bax, Bcl2, and cleaved caspase-3, as well as HSP70 and heat shock factor-1 (HSF1), were measured by Western blot. The melatonin effect through HSP70 was tested with silencing of HSF1 in the MPP + -treated SH-SY5Y cells. Melatonin can protect against MPP + -induced neuronal toxicity by promoting anti-oxidative and anti-apoptotic properties. SH-SY5Y cells exposed to melatonin with MPP + showed increased expression of HSP70 and HSF1 compared with those exposed to MPP + alone. However, siRNA-mediated downregulation of HSF1 significantly attenuated the protective effects of melatonin in the MPP + -induced in vitro PD model. Our findings revealed the protective roles of melatonin in an in vitro PD model. Melatonin can hinder the toxic effects of MPP + on dopaminergic neuronal cells via upregulation of the HSF1/HSP70 pathway. Further experimental studies would verify the therapeutic relevance of melatonin with HSP70 and HSF1 to prevent and decelerate PD-like neurodegeneration.
热休克蛋白 (HSPs) 作为分子伴侣发挥着重要作用，可以防止异常蛋白聚集和错误折叠。此外，它们还可以保护多巴胺神经元免受氧化应激、炎症和细胞凋亡的影响，这些都是帕金森病 (PD) 的众所周知的发病机制。褪黑素是一种有效的抗氧化剂，具有预防 PD 等神经退行性疾病的有益能力。我们旨在探索褪黑素在体外 PD 模型中的保护特性，重点研究 HSPs 的潜在机制。使用维甲酸 (RA) 分化的 SH-SY5Y 细胞建立 1-甲基-4-苯基吡啶鎓 (MPP+) 诱导的毒素模型。使用 MTT 和 DAPI 测量细胞活力和细胞凋亡。通过细胞通透性荧光探针 DCFH-DA 测量细胞内活性氧 (ROS) 水平。使用 ELISA 试剂盒评估丙二醛水平以及超氧化物歧化酶和谷胱甘肽过氧化物酶的活性。通过 Western blot 测量 Bax、Bcl2 和 cleaved caspase-3 以及 HSP70 和热休克因子-1 (HSF1) 的凋亡标志物。通过在 MPP+处理的 SH-SY5Y 细胞中沉默 HSF1 测试褪黑素对 HSP70 的作用。褪黑素可以通过促进抗氧化和抗凋亡特性来保护神经元免受 MPP+诱导的神经毒性。与单独暴露于 MPP+的细胞相比，暴露于 MPP+和褪黑素的 SH-SY5Y 细胞显示 HSP70 和 HSF1 的表达增加。然而，siRNA 介导的 HSF1 下调显著减弱了褪黑素在体外 PD 模型中的保护作用。我们的研究结果揭示了褪黑素在体外 PD 模型中的保护作用。褪黑素可以通过上调 HSF1/HSP70 通路来阻止 MPP+对多巴胺能神经元细胞的毒性作用。进一步的实验研究将验证 HSP70 和 HSF1 与褪黑素在预防和减缓 PD 样神经退行性变方面的治疗相关性。
