脊髓损伤再生机制及人神经干细胞分泌组治疗在大鼠模型中的作用
Mechanism of spinal cord injury regeneration and the effect of human neural stem cells-secretome treatment in rat model.
作者信息
Semita I Nyoman, Utomo Dwikora Novembri, Suroto Heri
机构信息
Doctoral Program of Medical Science, Faculty of Medicine, Universitas Airlangga, Surabaya 60132, Indonesia.
Department of Orthopedic and Traumatology, Faculty of Medicine, University of Jember, Jember 68121, Indonesia.
出版信息
World J Orthop. 2023 Feb 18;14(2):64-82. doi: 10.5312/wjo.v14.i2.64.
BACKGROUND
Globally, complete neurological recovery of spinal cord injury (SCI) is still less than 1%, and 90% experience permanent disability. The key issue is that a pharmacological neuroprotective-neuroregenerative agent and SCI regeneration mechanism have not been found. The secretomes of stem cell are an emerging neurotrophic agent, but the effect of human neural stem cells (HNSCs) secretome on SCI is still unclear.
AIM
To investigate the regeneration mechanism of SCI and neuroprotective-neuroregenerative effects of HNSCs-secretome on subacute SCI post-laminectomy in rats.
METHODS
An experimental study was conducted with 45 Rattus norvegicus, divided into 15 normal, 15 control (10 mL physiologic saline), and 15 treatment (30 μL HNSCs-secretome, intrathecal T10, three days post-traumatic). Locomotor function was evaluated weekly by blinded evaluators. Fifty-six days post-injury, specimens were collected, and spinal cord lesion, free radical oxidative stress (F2-Isoprostanes), nuclear factor-kappa B (NF-κB), matrix metallopeptidase 9 (MMP9), tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10), transforming growth factor-beta (TGF-β), vascular endothelial growth factor (VEGF), B cell lymphoma-2 (Bcl-2), nestin, brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF) were analyzed. The SCI regeneration mechanism was analyzed using partial least squares structural equation modeling (PLS SEM).
RESULTS
HNSCs-secretome significantly improved locomotor recovery according to Basso, Beattie, Bresnahan (BBB) scores and increased neurogenesis (nestin, BDNF, and GDNF), neuroangiogenesis (VEGF), anti-apoptotic (Bcl-2), anti-inflammatory (IL-10 and TGF-β), but decreased pro-inflammatory (NF-κB, MMP9, TNF-α), F2-Isoprostanes, and spinal cord lesion size. The SCI regeneration mechanism is valid by analyzed outer model, inner model, and hypothesis testing in PLS SEM, started with pro-inflammation followed by anti-inflammation, anti-apoptotic, neuroangiogenesis, neurogenesis, and locomotor function.
CONCLUSION
HNSCs-secretome as a potential neuroprotective-neuroregenerative agent for the treatment of SCI and uncover the SCI regeneration mechanism.
背景
在全球范围内，脊髓损伤（SCI）后神经功能完全恢复的比例仍低于1%，90%的患者会出现永久性残疾。关键问题在于尚未找到具有神经保护 - 神经再生作用的药物及SCI的再生机制。干细胞分泌组是一种新兴的神经营养因子，但人神经干细胞（HNSCs）分泌组对SCI的影响仍不清楚。
目的
探讨SCI的再生机制以及HNSCs分泌组对大鼠椎板切除术后亚急性SCI的神经保护 - 神经再生作用。
方法
以45只大鼠进行实验研究，分为15只正常组、15只对照组（10 mL生理盐水）和15只治疗组（创伤后3天，鞘内注射30 μL HNSCs分泌组于T10）。由不知情的评估者每周评估运动功能。损伤后56天，收集标本，分析脊髓损伤、自由基氧化应激（F2 - 异前列腺素）、核因子 - κB（NF - κB）、基质金属蛋白酶9（MMP9）、肿瘤坏死因子 - α（TNF - α）、白细胞介素 - 10（IL - 10）、转化生长因子 - β（TGF - β）、血管内皮生长因子（VEGF）、B细胞淋巴瘤 - 2（Bcl - 2）、巢蛋白、脑源性神经营养因子（BDNF）、胶质细胞源性神经营养因子（GDNF）。使用偏最小二乘结构方程模型（PLS SEM）分析SCI的再生机制。
结果
根据Basso、Beattie、Bresnahan（BBB）评分，HNSCs分泌组显著改善了运动功能恢复，并增加了神经发生（巢蛋白、BDNF和GDNF）、神经血管生成（VEGF）、抗凋亡（Bcl - 2）、抗炎（IL - 10和TGF - β），但降低了促炎（NF - κB、MMP9、TNF - α）、F2 - 异前列腺素和脊髓损伤大小。通过PLS SEM中的外部模型、内部模型和假设检验分析，SCI的再生机制是有效的，始于促炎，随后是抗炎、抗凋亡、神经血管生成、神经发生和运动功能。
结论
HNSCs分泌组作为一种潜在的神经保护 - 神经再生剂可用于治疗SCI，并揭示了SCI的再生机制。
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