目的 探讨嗅黏膜神经干细胞(OM-NSCs)对大鼠神经元凋亡的影响。方法 从雄性Sprague-Dawley大鼠分离提取OM-NSCs,从Sprague-Dawley胎鼠分离神经元。体外培养后,神经元分为空白组、对照组和实验组。空白组正常培养,对照组加白细胞介素-1β (IL-1β)刺激24 h后正常培养,实验组于IL-1β刺激后加OM-NSCs共培养。24 h后使用TUNEL和微管结合蛋白2 (MAP2)抗体双标染色观察神经元凋亡。结果 空白组仅有少量细胞凋亡,对照组出现大量凋亡细胞,实验组凋亡细胞数较对照组明显减少(F=39.764, P<0.01)。结论 OM-NSCs能抑制大鼠神经元凋亡,具有神经保护作用。
Objective To investigate the effect of olfactory mucosal neural stem cells (OM-NSCs) on neuronal apoptosis in rats. Methods OM-NSCs were isolated from adult male Sprague-Dawley rats, and neurons were isolated from fetal Sprague-Dawley rats. The neurons were divided into blank group, control group and experiment group after culture in vitro, which were further cultured in nomal, cultured in nomal 24 hours after stimulation of interleukin-1β (IL-1β), and cultured with OM-NSCs 24 hours after stimulation of IL-1β, respectively, for 24 hours. The apoptosis of neurons was observed with double staining of TUNEL and microtubule-associated protein 2 antibodies. Results There were few apoptotic cells in the blank group, and very many in the control group; the apoptotic cells were fewer in the experiment group than in the control group (F=39.764, P<0.01).Conclusion OM-NSCs can significantly inhibit neuronal apoptosis, which may play a role in neuroprotection.
[1] van der Scheer JW, Hutchinson MJ, Paulson T, et al. Reliability and validity of subjective measures of aerobic intensity in adults with spinal cord injury: a systematic review[J]. PM R, 2018, 10(2): 194-207.
[2] Biglari B, Swing T, Child C, et al.A pilot study on temporal changes in IL-1β and TNF-α serum levels after spinal cord injury: the serum level of TNF-α in acute SCI patients as a possible marker for neurological remission[J]. Spinal Cord, 2015, 53(7): 510-514.
[3] Curtis E, Martin JR, 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.
[4] Carelli S, Giallongo T, Gerace C, et al.Neural stem cell transplantation in experimental contusive model of spinal cord injury[J]. J Vis Exp, 2014, (94). doi: 10.3791/52141.
[5] Yao R, Murtaza M, Velasquez JT, et al.Olfactory ensheathing cells for spinal cord injury: sniffing out the issues[J]. Cell Transplant, 2018, 27(6): 879-889.
[6] Gou Y, Liu D, Liu J, et al.[Protective effect of?transplantation?of bone mesenchymal?stem?cells?on demyelination in?spinal?cord injury][J]. [in Chinese]. Sheng Wu Gong Cheng Xue Bao, 2018, 34(5): 761-776.
[7] Boese AC, Le QE, Pham D, et al.Neural stem cell?therapy for subacute and chronic ischemic stroke[J]. Stem Cell Res Ther, 2018, 9(1): 154.
[8] Bonner JF, Steward O.Repair of spinal cord injury with neuronal relays: from fetal grafts to neural stem cells[J]. Brain Res, 2015, 1619: 115-123.
[9] Xiao Z, Tang F, Zhao Y, et al.Significant improvement of acute complete?spinal?cord injury patients diagnosed by a combined criteria implanted with NeuroRegen Scaffolds and mesenchymal?stem?cells[J]. Cell Transplant, 2018, 27(6): 907-915.
[10] Miura T, Kikuchi N, Ohtsuka M, et al.Varicella in a patient with drug-induced hypersensitivity syndrome developed after autologous peripheral stem cell transplantation[J]. Int J Dermatol, 2018. doi: 10.1111/ijd.14091.[Epub ahead of print]
[11] Watzlawick R, Rind J, Sena ES, et al.Olfactory ensheathing cell transplantation in experimental spinal cord injury: effect size and reporting bias of 62 experimental treatments: a systematic review and meta-analysis[J]. PLoS Biol, 2016, 14(5): e1002468.
[12] Zhuo Y, Wang L, Ge L, et al.Hypoxic culture promotes dopaminergic-neuronal differentiation of nasal olfactory mucosa mesenchymal stem cells via upregulation of hypoxia-inducible factor-1α[J]. Cell Transplant, 2017, 26(8): 1452-1461.
[13] Stamegna JC, Girard SD, Veron A, et al.A unique method for the isolation of nasal olfactory stem cells in living rats[J]. Stem?Cell Res, 2014, 12(3): 673-679.
[14] 王超,崔国兴,王国祥,等. 嗅黏膜神经干细胞的体外培养和向酪氨酸羟化酶阳性神经元的分化[J]. 神经解剖学杂志, 2010, 26(4): 360-366.
[15] 谢扬智,李乾露,杨德雨,等. 人嗅黏膜神经干细胞体外培养及氟西汀对其增殖和神经发生的影响[J]. 中国细胞生物学学报, 2016, 38(12): 1480-1486.
[16] Kwon BK, Sekhon LH, Fehlings MG.Emerging repair, regeneration, and translational research advances for spinal cord injury[J]. Spine (Phila Pa 1976), 2010, 35(21 Suppl): S263-S270.
[17] Sutherland TC, Mathews KJ, Mao Y, et al.Differences in the cellular response to acute spinal cord injury between developing and mature rats highlights the potential significance of the inflammatory response[J]. Front Cell Neurosci, 2017, 10: 310.
[18] Nicola F, Marques MR, Odorcyk F, et al.Stem?cells?from human exfoliated deciduous teeth modulate early astrocyte response after?spinal?cord contusion[J]. Mol Neurobiol, 2018. doi: 10.1007/s12035-018-1127-4.[Epub ahead of print]
[19] McMahill BG, Spriet M, Sisó S, et al. Feasibility study of canine epidermal?neural?crest?stem?cell transplantation in the?spinal?cords of dogs[J]. Stem?Cells?Transl Med, 2015, 4(10): 1173-1186.
[20] Xiong LL, Zou Y, Shi Y, et al.Tree shrew?neural stem cell?transplantation?promotes functional recovery of tree shrews with a hemi sectioned?spinal?cord injury by upregulating nerve growth factor expression[J]. Int J Mol Med, 2018, 41(6): 3267-3277.
[21] Matsushita T, Lankford KL, Arroyo EJ, et al.Diffuse and persistent blood-spinal?cord?barrier disruption after contusive?spinal cord injury?rapidly recovers following intravenous infusion of?bone marrow?mesenchymal stem cells[J]. Exp Neurol, 2015, 267: 152-164.
[22] Gómez RM, Sánchez MY, Portela-Lomba M, et al.Cell therapy for spinal cord injury with olfactory ensheathing glia cells (OECs)[J]. Glia, 2018, 66(7): 1267-1301.
[23] Bagher Z, Kamrava SK, Alizadeh R, et al.Differentiation of neural crest?stem cells?from nasal mucosa into motor neuron-like?cells[J]. J Chem Neuroanat, 2018, 92: 35-40.
[24] Ueha R, Shichino S, Ueha S, et al.Reduction of proliferating?olfactory?cells?and low expression of extracellular matrix genes are hallmarks of the aged?olfactory?mucosa[J]. Front Aging Neurosci, 2018, 10: 86.
[25] Zhou R, Xu T, Liu X, et al.Activation of?spinal?dorsal horn P2Y13?receptors can promote the expression of IL-1β and?IL-6?in rats with diabetic neuropathic pain[J]. J Pain Res, 2018, 11: 615-628.
[26] Yang X, Chen S, Shao Z, et al.Apolipoprotein E deficiency exacerbates?spinal cord injury?in mice: inflammatory response and oxidative stress mediated by NF-κB signaling pathway[J]. Front Cell Neurosci, 2018, 12: 142.
