Institute of Basic Medical Sciences & School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China.
Toxicology. 2011 Jan 11;279(1-3):189-95. doi: 10.1016/j.tox.2010.10.011. Epub 2010 Oct 30.
Glutamate excitotoxicity has been implicated as one of the factors contributing to neuronal apoptosis and is involved in many neurodegenerative diseases. Previous studies suggest that mesenchymal stem cells have the ability to protect cultured neurons from excitotoxicity-induced apoptosis, although the underlying mechanisms are not clear. In this study, we evaluated whether adipose mesenchymal stem cells (AMSCs) could protect against glutamate-induced injury in PC12 cells by secreting neurotrophic factors. We found that AMSCs secreted neurotrophic factors including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) under both normoxic and hypoxic conditions. AMSC - conditioned medium (AMSC-CM) had a protective effect on excitotoxicity-injured PC12 cells, as indicated by increased cell viability, decreased number of TUNEL-staining positive nuclei and lowered caspase-3 activity. By using neutralizing monoclonal antibodies and specific inhibitors, VEGF, HGF and BDNF were identified as the mediators of AMSC effects and PI3-K/Akt and MAPK pathways were involved. Western blot analysis showed that AMSC-CM can increase the level of p-Akt, up-regulate XIAP and reduce the level of cleaved-caspase-3 in PC12 cells. These results suggest that AMSCs can effectively protect PC12 cells from glutamate excitotoxicity-induced apoptosis and support the hypothesis that AMSCs may be a useful treatment for stroke or neurodegenerative diseases which often involve excitotoxicity.
谷氨酸兴奋性毒性已被认为是导致神经元凋亡的因素之一,并且与许多神经退行性疾病有关。先前的研究表明,间充质干细胞具有保护培养神经元免受兴奋性毒性诱导的细胞凋亡的能力,尽管其潜在机制尚不清楚。在这项研究中,我们评估了脂肪间充质干细胞(AMSCs)是否可以通过分泌神经营养因子来防止谷氨酸诱导的 PC12 细胞损伤。我们发现,在常氧和低氧条件下,AMSCs 分泌包括血管内皮生长因子(VEGF)、肝细胞生长因子(HGF)、脑源性神经营养因子(BDNF)和神经生长因子(NGF)在内的神经营养因子。AMSC 条件培养基(AMSC-CM)对兴奋性毒性损伤的 PC12 细胞具有保护作用,表现为细胞活力增加、TUNEL 染色阳性核数量减少和 caspase-3 活性降低。通过使用中和单克隆抗体和特异性抑制剂,鉴定出 VEGF、HGF 和 BDNF 是 AMSC 作用的介导物,并且涉及 PI3-K/Akt 和 MAPK 途径。Western blot 分析表明,AMSC-CM 可以增加 PC12 细胞中 p-Akt 的水平,上调 XIAP 并降低 cleaved-caspase-3 的水平。这些结果表明,AMSCs 可以有效保护 PC12 细胞免受谷氨酸兴奋性毒性诱导的细胞凋亡,并支持 AMSCs 可能是治疗中风或神经退行性疾病的有用方法的假说,这些疾病通常涉及兴奋性毒性。