【摘要】：目的:評(píng)估低氧預(yù)處理臍帶間充質(zhì)干細(xì)胞(human umbilical cord mesenchymal stem cells,UCMSCs)移植修復(fù)大鼠急性脊髓損傷的作用并初步探討其機(jī)制。方法:分別于常氧和低氧條件下培養(yǎng)UCMSCs,細(xì)胞免疫熒光鑒定細(xì)胞,ELISA測(cè)定細(xì)胞腦源性生長(zhǎng)因子(BDNF)、血管源性生長(zhǎng)因子(VEGF)、睫狀生長(zhǎng)因子(CNTF)和肝細(xì)胞生長(zhǎng)因子(HGF)的濃度。采用改良的Al1en’s法(2.5 g×10 cm)建立60只大鼠T10脊髓損傷模型,隨機(jī)分為假手術(shù)組、模型組、低氧細(xì)胞移植組及常氧細(xì)胞移植組,每組20只,損傷后即刻移植1×10~6個(gè)細(xì)胞至低氧細(xì)胞移植組及常氧細(xì)胞移植組動(dòng)物脊髓內(nèi)。術(shù)前及術(shù)后每周進(jìn)行運(yùn)動(dòng)功能BBB評(píng)分;采用免疫熒光法檢測(cè)移植細(xì)胞的存活、神經(jīng)細(xì)胞凋亡;采用ELISA法檢測(cè)細(xì)胞移植第2、14、28 d脊髓組織肝細(xì)胞生長(zhǎng)因子的濃度。結(jié)果:在低氧組,體外由UCMSCs分泌的BDNF、VEGF、CNTF和HGF濃度顯著高于常氧組細(xì)胞。與模型組比較,細(xì)胞移植組運(yùn)動(dòng)功能評(píng)分和HGF含量增加,而神經(jīng)細(xì)胞的凋亡率降低,但以低氧細(xì)胞移植組改變最明顯。結(jié)論:低氧預(yù)處理的UMCSCs具有提高神經(jīng)營(yíng)養(yǎng)因子分泌作用,其機(jī)制可能與下調(diào)神經(jīng)細(xì)胞凋亡,增加受損脊髓組織HGF有關(guān)。
[Abstract]:Aim: to evaluate the effect of hypoxic preconditioning (human umbilical cord mesenchymal stem cells,UCMSCs) transplantation on the repair of acute spinal cord injury (sci) in rats. Methods: UCMSCs, cells were cultured under normoxic and hypoxic conditions, respectively. The concentrations of (BDNF), (VEGF), ciliary growth factor (CNTF) and hepatocyte growth factor (HGF) were determined by Elisa. 60 T10 spinal cord injury models were established by modified Al1en's method (2.5 g 脳 10 cm) and were randomly divided into sham-operation group, model group, hypoxic cell transplantation group and normoxic cell transplantation group, with 20 rats in each group. Immediately after injury, 1 脳 10 ~ 6 cells were transplanted into the spinal cord of rats in hypoxic cell transplantation group and normoxic cell transplantation group. BBB score of motor function was performed before and every week after transplantation; survival of transplanted cells and apoptosis of nerve cells were detected by immunofluorescence method; and the concentration of hepatocyte growth factor in spinal cord tissue was detected by ELISA method on the 28th day after transplantation. Results: the concentrations of BDNF,VEGF,CNTF and HGF secreted by UCMSCs in hypoxia group were significantly higher than those in normoxic group. Compared with the model group, the motor function score and HGF content in the transplantation group increased, while the apoptosis rate of the nerve cells decreased, but the changes were most obvious in the hypoxic cell transplantation group. Conclusion: hypoxia preconditioning UMCSCs can increase the secretion of neurotrophic factor, and its mechanism may be related to down-regulation of neuronal apoptosis and increase of HGF in injured spinal cord tissue.
【作者單位】： 南方醫(yī)科大學(xué)珠江醫(yī)院骨科;
【基金】：國(guó)家自然科學(xué)基金-青年科學(xué)基金(81400996) 廣東省自然科學(xué)基金-博士啟動(dòng)項(xiàng)目(2014A030310100)
【分類號(hào)】：R651.2

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