A novel cytoprotective peptide protects mesenchymal stem cells against mitochondrial dysfunction and apoptosis induced by starvation via Nrf2/Sirt3/FoxO3a pathway.
A novel cytoprotective peptide protects mesenchymal stem cells against mitochondrial dysfunction and apoptosis induced by starvation via Nrf2/Sirt3/FoxO3a pathway.
复制标题
一种新型细胞保护肽通过 Nrf2/Sirt3/FoxO3a 途径保护间充质干细胞免受饥饿诱导的线粒体功能障碍和细胞凋亡
DOI：
10.1186/s12967-017-1144-5
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发表时间：
2017-02-15
影响因子：
7.4
通讯作者：
Zhu T
中科院分区：
文献类型：
--
作者：
Wang S;Zhang C;Niyazi S;Zheng L;Li J;Zhang W;Xu M;Rong R;Yang C;Zhu T
背景间充质干细胞（mesenchymal stem cell，MSC）作为一种细胞治疗方法在过去的十年中得到了广泛的研究。然而，适应性转移的MSC的存活率差限制了其临床治疗潜力，这在很大程度上归因于营养饥饿。在这项研究中，我们确定了一种新的肾脏保护肽CHBP是否可以保护骨髓间充质干细胞免受饥饿和投资的潜在mechanism.MethodsMSCs进行血清剥夺和CHBP的梯度浓度管理。CCK-8法、Annexin V/PI法和Hoechst染色法检测细胞活力和凋亡。用流式细胞术测定ROS产生、JC-1指示的线粒体膜电位和线粒体质量。荧光显微镜下观察细胞色素c在细胞内的定位。Western blot分析Nrf 2、Sirt 3和FoxO 3a的表达。此外，应用CHBP预处理MSC以测试可能的抗饥饿保护作用。最后，CHBP对骨髓间充质干细胞的分化和自我更新能力的影响也examined.ResultsCHBP提高细胞活力和抑制凋亡的剂量依赖性方式。饥饿可导致线粒体功能障碍，CHBP可通过减少ROS的氧化损伤，恢复线粒体膜电位，维持线粒体膜的完整性，减轻线粒体应激。更重要的是，CHBP激活了Nrf 2/Sirt 3/FoxO 3a通路，并且Sirt 3敲低部分取消了CHBP的保护作用。此外，CHBP预处理的MSC更耐饥饿。在正常条件下，CHBP对MSCs的分化和自我更新能力影响不大。ConclusionsThe目前的研究表明，CHBP对饥饿诱导的MSCs线粒体功能障碍和凋亡具有有效的保护作用，并表明Nrf 2/Sirt 3/FoxO 3a通路可能参与了这种保护作用。
BackgroundMesenchymal stem cell (MSC) has been widely explored in the past decade as a cell-based treatment for various diseases. However, poor survival of adaptively transferred MSCs limits their clinical therapeutic potentials, which is largely ascribed to the nutrient starvation. In this study, we determined whether a novel kidney protective peptide CHBP could protect MSCs against starvation and invested the underlying mechanisms.MethodsMSCs were subjected to serum deprivation and CHBP of graded concentrations was administered. Cell viability and apoptosis were detected by CCK-8, Annexin V/PI assay and Hoechst staining. ROS generation, mitochondrial membrane potential indicated by JC-1 and mitochondrial mass were measured by flow cytometry. The location of cytochrome c within cells was observed under fluorescence microscopy. Expressions of Nrf2, Sirt3, and FoxO3a were analyzed by western blot. In addition, preconditioning MSCs with CHBP was applied to test the possible protection against starvation. Finally, the effect of CHBP on the differentiation and self-renewal capacity of MSCs was also examined.ResultsCHBP improved cell viability and suppressed apoptosis in a dose dependent manner. Starvation resulted in the mitochondrial dysfunction and treatment of CHBP could alleviate mitochondrial stress by diminishing oxidative injury of ROS, restoring mitochondrial membrane potential and maintaining mitochondrial membrane integrity. Importantly, Nrf2/Sirt3/FoxO3a pathway was activated by CHBP and Sirt3 knockdown partially abolished the protection of CHBP. Moreover, MSCs pretreated with CHBP were more resistant to starvation. Under normal condition, CHBP exerted little effects on the differential and self-renewal capacity of MSCs.ConclusionsThe present study demonstrated the efficient protection of CHBP upon MSCs against starvation-induced mitochondrial dysfunction and apoptosis and indicated possible involvement of Nrf2/Sirt3/FoxO3a pathway in the protective effect.