Li Zhi, Wu Haihui
Department of Orthopedics, Qingpu Branch of Zhongshan Hospital, Fudan University, 201700, P.R.China.
Department of Orthopedics, Qingpu Branch of Zhongshan Hospital, Fudan University, 201700,
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2017 Nov 15;31(11):1377-1383. doi: 10.7507/1002-1892.201706082.
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.
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 ( =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.
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 ( <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 ( <0.05); but there was no significant difference between groups C and D ( >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 ( <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 ( <0.05).
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.
探讨人尿源干细胞（hUSCs）及hUSCs联合软骨素酶ABC（chABC）对大鼠脊髓损伤（SCI）后神经生长因子（NGF）和脑源性神经营养因子（BDNF）表达的影响，并探究其潜在机制。
从人尿中培养hUSCs，通过流式细胞术检测其表型。采用Allen法制备大鼠SCI模型。将60只Sprague Dawley大鼠分为5组（每组n = 12）：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）。于术后10、20和30天，采用Basso、Beattie、Bresnahan（BBB）评分法评估各组大鼠下肢运动功能。采用实时荧光定量PCR检测术后30天NGF和BDNF的相对mRNA表达。同时，通过免疫组织化学染色确认NGF和BDNF的蛋白表达。采用蛋白质印迹法检测Bax和Bcl-2的相对蛋白表达。
鉴定hUSCs具有多向分化潜能。术后10、20和30天，B组BBB评分显著低于A、C、D、E组，C、D、E组低于A组，C、D组低于E组（P<0.05）。实时荧光定量PCR和免疫组织化学染色显示，B组NGF和BDNF表达显著低于A、C、D、E组，C、D、E组低于A组，C、D组低于E组（P<0.05）；但C、D组间差异无统计学意义（P>0.05）。蛋白质印迹结果表明，B组Bax蛋白表达显著高于A、C、D、E组，C、D、E组高于A组，C、D组高于E组（P<0.05）。同时，B组Bcl-2蛋白表达显著低于A、C、D、E组，C、D、E组低于A组，C、D组低于E组（P<0.05）。
hUSCs可对SCI起到保护作用，chABC可增强这种积极作用；这种神经保护作用可能依赖于促进NGF和BDNF的表达以及抑制神经元凋亡。
