目的：观察高温预处理嗅鞘细胞（olfactory ensheathing cells,OECs）联合神经干细胞（neural stem cells,NSCs）修复脊髓损伤的效果,为临床治疗脊髓损伤提供参考。方法：分别采用普通与40℃预处理的OECs上清液对NSCs进行培养,比较两组NSCs向神经元分化情况;建立脊髓半横断损伤模型,分别采用普通OECs、高温预处理OECs与NSCs联合移植治疗脊髓损伤大鼠,对照组注射PBS,观察大鼠运动恢复情况及脊髓病理改变,分析高温预处理OECs对脊髓损伤修复的影响。结果：细胞实验中,高温预处理组NSCs向神经元分化比例明显高于对照组。动物实验中,高温预处理组大鼠运动功能BBB评分在移植后2周后快速上升,6周后进入缓慢上升阶段,增长幅度与速度明显高于另外两组,运动功能恢复明显,爬杆评分和旋转时间结果与BBB评分类似。脊髓病理切片显示,高温预处理组脊髓组织结构恢复效果最佳,BrdU示踪染色显示移植细胞广泛分布于修复后损伤区域。结论：高温预处理OECs可促进NSCs向神经元分化,两者联合移植可有效治疗脊髓损伤。
Abstract
Objective To observe the effect of Hyperthermia preconditioning olfactory ensheathing cells (OECs) combined with neural stem cells (NSCs) on spinal cord injury, and to provide reference for clinical treatment of spinal cord injury. Methods NSCs were cultured with normal or 40℃ pretreated OECs supernatant respectively, and the number of NSCs differentiation into neurons were compared between the two groups. The spinal cord semi-transection injury model was established, then the rata with spinal cord injury were treated by combined transplantation of NSCs with OECs or hyperthermia pretreated OECs . Control group was injected with PBS. The motor recovery and pathological changes of the rats were observed, and the effect of hyperthermia preconditioning OECs on the repair of spinal cord injury was analyzed. Results In the cell experiments, the proportion of NSCs differentiation into neurons in the hyperthermia preconditioning group was significantly higher than that in the control group. In animal experiments, the BBB scores of motor function in the hyperthermia preconditioning group increased rapidly 2 weeks after transplantation, and entered the slow rising stage 6 weeks after transplantation. The increase amplitude and speed were significantly higher than those in the other two groups, and the motor function recovered obviously. The results of rod climbing score and rotation time were similar to the BBB score. The pathological section of spinal cord showed that the hyperthermia preconditioning group had the best recovery effect of spinal cord tissue structure. BrdU tracer staining showed that the transplanted cells were widely distributed in the damaged area after repair. Conclusion Hyperthermia preconditioning of OECs can promote the differentiation of NSCs into neurons. Combined transplantation of Hyperthermia preconditioning OECs and NSCs can effectively treat spinal cord injury.
关键词
高温预处理 /
脊髓损伤 /
嗅鞘细胞 /
神经干细胞
Key words
hyperthermia preconditioning /
spinal cord injury /
olfactory ensheathing cells /
neural stem cells
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] Kadoya K, Lu P, Nguyen K, et al.Spinal cord reconstitution with homologous neural grafts enables robust corticospinal regeneration[J]. Natu Med, 2016, 22(5): 479-487.
[2] Zhou P, Guan J, Xu P, et al.Cell Therapeutic Strategies for Spinal Cord Injury[J]. Adv Wound Care (New Rochelle), 2019, 8(11): 585-605.
[3] Lu P, Ceto S, Wang Y, et al.Prolonged human neural stem cell maturation supports recovery in injured rodent CNS[J]. J Clin Invest, 2017, 127(9): 3287-3299.
[4] Wang G, Ao Q, Gong K, et al.Synergistic effect of neural stem cells and olfactory ensheathing cells on repair of adult rat spinal cord injury[J]. Cell Transplant, 2010, 19(10): 1325-1337.
[5] Shpargel KB, Jalabi W, Jin Y, et al. Preconditioning paradigms and pathways in the brain [J]. Cleve Clin J Med, 2008, 75 Suppl 2: S77-82.
[6] Chen C R, Li Y C, Young T H.Gallium nitride induces neuronal differentiation markers in neural stem/precursor cells derived from rat cerebral cortex[J]. Acta biomater, 2009, 5(7): 2610-2617.
[7] Nash H H, Borke R C, Anders J J.New method of purification for establishing primary cultures of ensheathing cells from the adult olfactory bulb[J]. Glia, 2001, 34(2): 81-87.
[8] Basso D M, Beattie M S, Bresnahan J C.Graded histological and locomotor outcomes after spinal cord contusion using the NYU weight-drop device versus transection[J]. Exp Neurol, 1996, 139(2): 244-256.
[9] Brailowsky S, Knight RT, Blood K, et al.γ-Aminobutyric acid-induced potentiation of cortical hemiplegia[J]. Brain Res, 1986, 362(2): 322-330.
[10] Kleiser B, Van Reempts J, Van Deuren B, et al.Favourable effect of flunarizine on the recovery from hemiparesis in rats with intracerebral hematomas[J]. Neurosci Lett, 1989, 103(2): 225-228.
[11] Curtis E, Martin J R, Gabel B, et al. A First-in-Human, Phase I Study of Neural Stem Cell Transplantation for Chronic Spinal Cord Injury [J]. Cell Stem Cell, 2018, 22(6): 941-950. e6.
[12] Xie J, Li Y, Dai J, et al.Olfactory Ensheathing Cells Grafted Into the Retina of RCS Rats Suppress Inflammation by Down-Regulating the JAK/STAT Pathway[J]. Front Cell Neurosci, 2019, 13: 341
[13] Li Y, Huo S, Fang Y, et al.ROCK Inhibitor Y27632 Induced Morphological Shift and Enhanced Neurite Outgrowth-Promoting Property of Olfactory Ensheathing Cells via YAP-Dependent Up-Regulation of L1-CAM[J]. Front Cell Neurosci, 2018, 12: 489.
[14] Wright AA, Todorovic M, Tello-Velasquez J, et al.Enhancing the Therapeutic Potential of Olfactory Ensheathing Cells in Spinal Cord Repair Using Neurotrophins[J]. Cell Transplant, 2018, 27(6): 867-878.
[15] Beckingham LJ, Todorovic M, Tello Velasquez J, et al.Three-dimensional cell culture can be regulated by vibration: low-frequency vibration increases the size of olfactory ensheathing cell spheroids[J]. J Biol Eng, 2019, 13: 41.
[16] Pespeni M, Hodnett M, Pittet JF.In vivo stress preconditioning[J]. Methods, 2005, 35(2): 158-164.
[17] Du F, Zhu L, Qian ZM, et al.Hyperthermic preconditioning protects astrocytes from ischemia/reperfusion injury by up-regulation of HIF-1 alpha expression and binding activity[J]. Biochim Biophys Acta, 2010, 1802(11): 1048-1053.
[18] Redaelli CA, Wagner M, Kulli C, et al.Hyperthermia-induced HSP expression correlates with improved rat renal isograft viability and survival in kidneys harvested from non-heart-beating donors[J]. Transpl Int, 2001, 14(6): 351-360.
[19] Xu H, Aibiki M, Nagoya J.Neuroprotective effects of hyperthermic preconditioning on infarcted volume after middle cerebral artery occlusion in rats: role of adenosine receptors[J]. Crit Care Med, 2002, 30(5): 1126-1130.
[20] Koffler J, Zhu W, Qu X, et al.Biomimetic 3D-printed scaffolds for spinal cord injury repair[J]. Nat Med, 2019, 25(2): 263-269.
基金
湖南省科技厅重点研发项目（2020SK2102）
PDF(4505 KB)
