Sun Xuejun, Fang Bo, Zhao Xi, Zhang Guangwei, Ma Hong
Department of Anesthesiology, First Affiliated Hospital, China Medical University, Shenyang, Liaoning, China.
Department of Cardiac Surgery, First Affiliated Hospital, China Medical University, Shenyang, Liaoning, China.
PLoS One. 2014 Mar 5;9(3):e90667. doi: 10.1371/journal.pone.0090667. eCollection 2014.
Bone marrow mesenchymal stem cells (MSCs) have been found to produce beneficial effects on ischemia-reperfusion injury. However, most of the MSCs died when transplanted into the ischemic tissue, which severely limit their therapeutic potential.
Using an in vitro model of hypoxia and serum deprivation (H/SD), we investigated the hypothesis that sevoflurane preconditioning could protect MSCs against H/SD-induced apoptosis and improve their migration, proliferation, and therapeutic potential. The H/SD of MSCs and neuron-like PC12 cells were incubated in a serum-free medium and an oxygen concentration below 0.1% for 24 h. Sevoflurane preconditioning was performed through a 2-h incubation of MSCs in an airtight chamber filled with 2 vol% sevoflurane. Apoptosis of MSCs or neuron-like PC12 cells was assessed using Annexin V-FITC/propidium iodide (PI). Furthermore, the mitochondrial membrane potential was assessed using lipophilic cationic probe. The proliferation rate was evaluated through cell cycle analysis. Finally, HIF-1α, HIF-2α, VEGF and p-Akt/Akt levels were measured by western blot.
Sevoflurane preconditioning minimized the MSCs apoptosis and loss of mitochondrial membrane potential. Furthermore, it increased the migration and expression of HIF-1α, HIF-2α, VEGF, and p-Akt/Akt, reduced by H/SD. In addition, neuron-like PC12 cells were more resistant to H/SD-induced apoptosis when they were co-cultured with sevoflurane preconditioning MSCs.
These findings suggest that sevoflurane preconditioning produces protective effects on survival and migration of MSCs against H/SD, as well as improving the therapeutic potential of MSCs. These beneficial effects might be mediated at least in part by upregulating HIF-1α, HIF-2α, VEGF, and p-Akt/Akt.
骨髓间充质干细胞（MSCs）已被发现对缺血再灌注损伤具有有益作用。然而，大多数MSCs在移植到缺血组织后死亡，这严重限制了它们的治疗潜力。
使用缺氧和血清剥夺（H/SD）的体外模型，我们研究了七氟醚预处理可以保护MSCs免受H/SD诱导的凋亡并改善其迁移、增殖和治疗潜力的假设。将MSCs和神经元样PC12细胞进行H/SD处理，即在无血清培养基和氧浓度低于0.1%的条件下孵育24小时。通过在充满2%七氟醚的密闭 chamber 中对MSCs进行2小时的孵育来进行七氟醚预处理。使用 Annexin V-FITC/碘化丙啶（PI）评估MSCs或神经元样PC12细胞的凋亡。此外，使用亲脂性阳离子探针评估线粒体膜电位。通过细胞周期分析评估增殖率。最后，通过蛋白质印迹法测量HIF-1α、HIF-2α、VEGF和p-Akt/Akt水平。
七氟醚预处理使MSCs凋亡和线粒体膜电位丧失最小化。此外，它增加了H/SD降低的HIF-1α、HIF-2α、VEGF和p-Akt/Akt的迁移和表达。此外，当与七氟醚预处理的MSCs共培养时，神经元样PC12细胞对H/SD诱导的凋亡更具抗性。
这些发现表明，七氟醚预处理对MSCs在H/SD条件下的存活和迁移产生保护作用，并提高了MSCs的治疗潜力。这些有益作用可能至少部分是通过上调HIF-1α、HIF-2α、VEGF和p-Akt/Akt介导的。
