【摘要】 背景:体内实验发现,骨髓间充质干细胞可在体内神经组织中分化为神经元样细胞,表达神经元抗原,并且骨髓间充质干细胞的存在对神经元的损伤有保护作用,这可能与骨髓间充质干细胞分泌的活性物质有关。目的:观察骨髓间充质干细胞移植联用诱导型一氧化氮合酶抑制剂-氨基胍对大鼠脊髓损伤修复的影响。设计、时间及地点:随机对照动物实验,细胞学观察,于2006-07/2008-02在河北工程大学附属医院外科实验室和中心实验室完成。材料:Wistar大鼠36只随机分成对照组、骨髓间充质干细胞组、联合移植组,每组12只。氨基胍由美国Sigma公司提供。方法:采用密度梯度离心法分离培养大鼠骨髓间充质干细胞,以Brdu标记细胞核。对照组:制备脊髓半切损伤模型不做处理。骨髓间充质干细胞组:制备脊髓半切损伤模型后,损伤上下各1mm处移植骨髓间充质干细胞悬液共2μL。联合移植组:制备脊髓半切损伤模型后,损伤上下各1mm处移植骨髓间充质干细胞悬液8μL,同时局部给予氨基胍100mg/kg。主要观察指标:术后采用斜板试验及改良Tarlov评分评价动物后肢运动功能恢复情况。术后2个月,显微镜下观察移植后脊髓形态结构变化及移植物存活情况。神经微丝和胶质原纤维酸性蛋白免疫组织化学评价移植对脊髓再生的影响。结果:下肢运动功能评价联合移植组优于骨髓间充质干细胞组,骨髓间充质干细胞组优于对照组。免疫组织化学检测发现,脊髓损伤区有新生束状轴突伸向断端,嗜银染色可见脊髓断端的再生轴突长入断端间组织中,部分连接两端面。损伤阶段脊髓内可见Brdu标记阳性骨髓间充质干细胞存活,灰质分布较白质多,以注射部位向周围损伤组织迁移,联合应用氨基胍组Brdu标记阳性骨髓间充质干细胞存活多。联合移植组术后2个月神经微丝和胶质原纤维酸性蛋白免疫阳性反应的面积比均高于其他各组。结论:骨髓间充质干细胞联合氨基胍移植促进大鼠半横断脊髓结构和功能恢复的效果明显优于单纯细胞移植组,两者联用具有协同效应。更多还原

【Abstract】 BACKGROUND: In vivo experiment showed that bone marrow mesenchymal stem cells (BMSCs) could differentiate into neuron-like cells and express neuron antigen in nervous tissue, and that the existence of BMSCs has protection on neuronal injury. It is probably concerned with active substance secreted by BMSCs. OBJECTIVE: To observe the effect of aminoguanidine combined with BMSC transplantation on recovery in rats after spinal cordinjury. DESIGN, TIME AND SETTING: Randomized, controlled animal cytology experiment was performed from July 2006 to February 2008 at the Surgery Laboratory and Central Laboratory in Hospital Affiliated to Hebei University of Engineering. MATERIALS: A total of 36 Wistar rats were divided into control, BMSCs, combination groups at random, 12 rats for each group. Aminoguanidine was supplied by Sigma, USA. METHODS: Rat BMSCs were obtained using density gradient centrifugation and nuclei were labeled by BrdU. Rat models of spinal cord hemisection were established in the control group. After establishment of rat models of spinal cord hemisection, totally2 μL BMSC suspension was injected into rats 1 mm up and down from injured region in the BMSC group. In the combination group, after establishment of rat models of spinal cord hemisection, totally 8 μL BMSC suspension was injected into rats 1 mm up and down from injured region, simultaneously, 100 mg/kg aminoguanidine was treated in local region. MAIN OUTCOME MEASURES: Locomotor function of animal hind limb was tested using the inclined plane test and modified Tarlov locomotor rating scale. At 2 months, morphous changes in spinal cord tissues and survival of transplant were observed with a microscopy. Immunohistochemical staining was conducted for neurofilament and glial fibrillary acidic protein to evaluate theeffect of transplantation on spinal cord regeneration. RESULTS: Motor function of hind limbs was better in the combination group than in the BMSC group, and better in the BMSC group than in the control group. Immunohistochemistry results showed that there was newly axis stretching to broken end of the spinal injury. The regenerated axis was observed in the edge of spinal cord injury using silver staining. The part of spinal cord injury presented Brdu-labeled BMSCs. There were more in grey matter than in white matter. It migrated from injection site to surrounding injured tissues. Brdu-labeled BMSCs were more in the combination group. Immunity positive reaction areas of neurofilament and glial fibrillary acidic protein survived more in the combination group compared with the BMSC group and the control group at two months after surgery. CONCLUSION: Aminoguanidine and BMSC transplantation can promote structure and functional rehabilitation of spinal cord injury. The outcome is significantly better than in the BMSC group. There may be a synergistic effect between aminoguanidine and BMSCs.更多还原