Neuroscience Research Group, Medical Research Institute, Faculty of Medicine, University of Antioquia (UdeA), SIU, Calle 70 No. 52-21, and Calle 62 # 52-59, Building 1, Room 412, Medellin, Colombia.
Mol Biol Rep. 2019 Aug;46(4):4423-4435. doi: 10.1007/s11033-019-04897-2. Epub 2019 May 30.
Parkinson disease (PD) is characterized by the loss of dopaminergic (DAergic) neurons linked to environmental toxicants that cause oxidative stress (OS). The aim of this investigation was to establish the molecular response of human mesenchymal stroma cells (MSCs) depleted of glutathione (GSH) by the specific inhibitor L-buthionine-sulfoximine (BSO) to 6-hydroxydopamine (6-OHDA) and/or N-acetylcysteine (NAC) co-treatment. We found that treatment with BSO (10 mM) plus 6-OHDA (200 μM) induced apoptosis in MSCs through an oxidative stress (OS) mechanism involving HO, reflected by the detection of dichlorofluorescein-positive (DCF+) cells and oxidation of DJ-1 Cys106-SH into DJ-1 Cys106-SO; an almost complete reduction in glutathione peroxidase 1 (GPX1) expression; activation of the transcription factor c-JUN, the pro-apoptotic protein BAX and BH-3-only protein PUMA; loss of mitochondrial membrane potential (∆Ψ); activation of the protease caspase-3 (CASP3) and apoptosis-inducing factor (AIF); chromatin condensation; and DNA fragmentation. Strikingly, co-treatment of MSCs with NAC (5 mM) and BSO + 6-OHDA significantly reduced the expression of OS and cell death markers but were unable to restore the expression of GPX1 compared to the expression in untreated or treated cells with NAC only. These findings highlighted the importance of the maintenance of the GSH-dependent (e.g., GPX1, GSH synthesis) and -independent (e.g., ROS scavenger molecules and thiol reducing activity) antioxidant systems (e.g., NAC) in the protection of MSCs from detrimental stress stimuli, thereby increasing the survival of stromal cells.
帕金森病（PD）的特征是多巴胺能（DAergic）神经元丧失，与导致氧化应激（OS）的环境毒物有关。本研究的目的是确定耗尽谷胱甘肽（GSH）的人间质基质细胞（MSCs）对 6-羟多巴胺（6-OHDA）和/或 N-乙酰半胱氨酸（NAC）共处理的分子反应。我们发现，用特异性抑制剂 L-丁硫氨酸亚砜（BSO）（10 mM）加 6-OHDA（200 μM）处理会通过涉及 HO 的氧化应激（OS）机制诱导 MSC 凋亡，这反映在检测到二氯荧光素阳性（DCF +）细胞和 DJ-1 Cys106-SH 氧化成 DJ-1 Cys106-SO 中；谷胱甘肽过氧化物酶 1（GPX1）的表达几乎完全降低；转录因子 c-JUN、促凋亡蛋白 BAX 和 BH-3 仅蛋白 PUMA 的激活；线粒体膜电位（∆Ψ）丧失；半胱天冬酶-3（CASP3）和凋亡诱导因子（AIF）的激活；染色质浓缩；和 DNA 片段化。引人注目的是，用 NAC（5 mM）和 BSO + 6-OHDA 共处理 MSC 可显著降低 OS 和细胞死亡标志物的表达，但与单独用 NAC 处理或未处理的细胞相比，GPX1 的表达无法恢复。这些发现强调了维持谷胱甘肽依赖性（例如，GPX1、GSH 合成）和非依赖性（例如，ROS 清除分子和硫醇还原活性）抗氧化系统（例如，NAC）在保护 MSC 免受有害应激刺激中的重要性，从而提高了基质细胞的存活率。
