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Title: [Effects of human urine-derived stem cells combined with chondroitinase ABC on the expressions of nerve growth factor and brain-derived neurotrophic factor in the spinal cord injury].
Author: Li Z, Wu H.
Journal: Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi; 2017 Nov 15; 31(11):1377-1383. PubMed ID: 29798595.
Abstract:
OBJECTIVE: To explore the effects of human urine-derived stem cells (hUSCs) and hUSCs combined with chondroitinase ABC (chABC) on the expressions of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in the spinal cord injury (SCI) of rats, and to investigate the underlying mechanism. METHODS: hUSCs were cultured from human urine, and their phenotypes were detected by flow cytometry. The SCI model of rats were made via Allen method. Sixty Sprague Dawley rats were divided into 5 groups ( n=12): the sham operation group (group A), SCI group (group B), SCI+hUSCs group (group C), SCI+chABC group (group D), and SCI+hUSCs+chABC group (group E). Basso, Beattie, Bresnahan (BBB) score was used to measure the lower extremity motor function of rats in each group at 10, 20, and 30 days after operation. Real-time fluorescent quantitative PCR was used to detect the relative mRNA expressions of NGF and BDNF at 30 days. Meanwhile, the protein expression of NGF and BDNF were confirmed by immunohistochemistry staining. The relative protein expressions of Bax and Bcl-2 were detected by Western blot. RESULTS: The hUSCs were identified to have multipotential differentiation potential. At 10, 20, and 30 days, BBB score was significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E ( P<0.05). Real-time fluorescent quantitative PCR and immunohistochemistry staining demonstrated that the expressions of NGF and BDNF were significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E ( P<0.05); but there was no significant difference between groups C and D ( P>0.05). Western blot results indicated that the protein expression of Bax was significantly higher in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E ( P<0.05). Meanwhile, the protein expression of Bcl-2 was significantly lower in group B than in groups A, C, D, and E, in groups C, D, and E than in group A, in groups C and D than in group E ( P<0.05). CONCLUSION: hUSCs can protect SCI and this positive effect can be enhanced by chABC; this neuro-protective effect may depend on promoting the expressions of NGF and BDNF, and suppressing the neuronal apoptosis. 目的: 探讨大鼠脊髓损伤（spinal cord injury，SCI）后，行人尿液干细胞（human urine-derived stem cells，hUSCs）移植以及联合硫酸软骨素酶 ABC（chondroitinase ABC，chABC）鞘内注射，对脊髓组织神经生长因子（nerve growth factor，NGF）及脑源性神经营养因子（brain-derived neurotrophic factor，BDNF）的影响，以及其相关分子机制。. 方法: 取人尿液分离培养 hUSCs 并鉴定。利用 Allen 法建立大鼠 SCI 模型；将 60 只大鼠随机分为 5 组（ n=12），分别为假手术组（A 组）、SCI 组（B 组）、SCI+hUSCs 组（C 组）、SCI+chABC 组（D 组）、SCI+hUSCs+chABC 组（E 组）。模型制备后 10、20、30 d，利用 BBB 评分法评价大鼠下肢运动功能；30 d 时取脊髓组织，采用实时荧光定量 PCR 检测及免疫组织化学染色观察 NGF、BDNF 表达情况，Western blot 检测 Bax、Bcl-2 蛋白表达。. 结果: 经鉴定，培养的细胞具有多向分化特性，为 hUSCs。SCI 模型制备后 10、20、30 d，B 组下肢运动功能 BBB 评分均显著低于 A、C、D、E 组，C、D、E 组显著低于 A 组，C、D 组低于 E 组，比较差异均有统计学意义（ P<0.05）。实时荧光定量 PCR 及免疫组织化学染色检测示，模型制备后 30 d，B 组 NGF、BDNF 表达显著低于 A、C、D、E 组，C、D、E 组显著低于 A 组，C、D 组低于 E 组，比较差异均有统计学意义（ P<0.05）；C、D 组间比较差异无统计学意义（ P>0.05）。Western blot 检测显示，B 组 Bax 蛋白相对表达量明显高于 A、C、D、E 组，C、D、E 组显著高于 A 组，C、D 组高于 E 组，比较差异均有统计学意义（ P<0.05）； B 组 Bcl-2 蛋白相对表达量显著低于 A、C、D、E 组，C、D、E 组显著低于 A 组，C、D 组低于 E 组，比较差异均有统计学意义（ P<0.05）。. 结论: hUSCs 对大鼠 SCI 具有修复作用，通过联合 chABC 可促进 SCI 修复，其机制可能是通过促进 NGF 和 BDNF 表达，抑制神经细胞凋亡，发挥保护作用。.

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