Mesenchymal Stem Cells (MSCs) Coculture Protects [Ca2+]i Orchestrated Oxidant Mediated Damage in Differentiated Neurons In Vitro
Mesenchymal Stem Cells (MSCs) Coculture Protects [Ca2+]i Orchestrated Oxidant Mediated Damage in Differentiated Neurons In Vitro
复制标题
间充质干细胞（MSCs）共培养可在体外保护分化神经元中由细胞内钙离子浓度（[Ca2+]i ）调控的、氧化应激介导的损伤 。
DOI：
10.3390/cells7120250
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发表时间：
2018-12-01
期刊：
影响因子：
6
通讯作者：
Chandramoorthy, Harish C.
中科院分区：
文献类型：
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作者：
Alhazzani, Adel;Rajagopalan, Prasanna;Chandramoorthy, Harish C.
由于间充质干细胞 (MSC) 在神经保护和再生中的作用，正在研究在实验性中风中使用间充质干细胞 (MSC) 的细胞治疗方式。有必要了解应激期间发生的事件顺序以及 MSC 如何补充 [Ca2+](i) 和活性氧 (ROS) 介导的神经元细胞死亡的救援。在目前的研究中，SH-SY5Y分化的神经元细胞在体外受到脑缺血样应激，并使用间充质干细胞/神经元细胞共培养模型实验性地将其从细胞死亡中拯救出来。神经元细胞死亡的特征是促炎性肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β和-12的诱导高达35倍，同时抗炎细胞因子转化生长因子（TGF）-β、IL-6和-10相应下调约1至7倍。细胞内钙 [Ca2+](i) 和 ROS 的增加清楚地证实了氧化应激介导的细胞凋亡，而核因子 NF-kappa B 和环加氧酶 (COX)-2 表达的上调，以及早期和晚期凋亡细胞类似 41% 的积累，证实了缺血应激介导的细胞死亡。当与 MSC 共培养时，通过增加抗炎细胞因子（TGF-β，17%；IL-6，4%；和 IL-10，13%）的表达，显着下调 NF-κ B 和促炎 COX-2 的表达，使应激神经元细胞免于死亡。早期和晚期凋亡细胞的进一步积累减少至23%，而相应的细胞死亡从40%减少至17%。 MSC 共培养可恢复 mRNA 水平的低超氧化物歧化酶 1 (SOD1) 表达，而过氧化氢酶 (CAT) 和谷胱甘肽过氧化物酶 (GPx) 则未观察到显着变化。有趣的是，培养上清液中血清素释放的增加与[Ca2+](i) 和相应ROS 的升高成正比，随后通过MSC 共培养将其恢复至接近正常状态。总而言之，所有这些结果主要支持 MSC 介导的体外应激神经元存活调节。
Cell-therapy modalities using mesenchymal stem (MSCs) in experimental strokes are being investigated due to the role of MSCs in neuroprotection and regeneration. It is necessary to know the sequence of events that occur during stress and how MSCs complement the rescue of neuronal cell death mediated by [Ca2+](i) and reactive oxygen species (ROS). In the current study, SH-SY5Y-differentiated neuronal cells were subjected to in vitro cerebral ischemia-like stress and were experimentally rescued from cell death using an MSCs/neuronal cell coculture model. Neuronal cell death was characterized by the induction of proinflammatory tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta and -12, up to 35-fold with corresponding downregulation of anti-inflammatory cytokine transforming growth factor (TGF)-beta, IL-6 and -10 by approximately 1 to 7 fold. Increased intracellular calcium [Ca2+](i) and ROS clearly reaffirmed oxidative stress-mediated apoptosis, while upregulation of nuclear factor NF-kappa B and cyclo-oxygenase (COX)-2 expressions, along with similar to 41% accumulation of early and late phase apoptotic cells, confirmed ischemic stress-mediated cell death. Stressed neuronal cells were rescued from death when cocultured with MSCs via increased expression of anti-inflammatory cytokines (TGF-beta, 17%; IL-6, 4%; and IL-10, 13%), significantly downregulated NF-kappa B and proinflammatory COX-2 expression. Further accumulation of early and late apoptotic cells was diminished to 23%, while corresponding cell death decreased from 40% to 17%. Low superoxide dismutase 1 (SOD1) expression at the mRNA level was rescued by MSCs coculture, while no significant changes were observed with catalase (CAT) and glutathione peroxidase (GPx). Interestingly, increased serotonin release into the culture supernatant was proportionate to the elevated [Ca2+](i) and corresponding ROS, which were later rescued by the MSCs coculture to near normalcy. Taken together, all of these results primarily support MSCs-mediated modulation of stressed neuronal cell survival in vitro.