摘要
背景:研究证实,Rho激酶抑制剂法舒地尔可以有效阻断继发性脊髓损伤过程。目的:观察法舒地尔对骨髓间充质干细胞移植治疗脊髓损伤大鼠运动功能恢复的影响。设计、时间及地点:随机对照动物实验,于2008-11/2009-03在天津医科大学内分泌研究所完成。材料:1月龄SD大鼠1只,用于分离制备骨髓间充质干细胞。健康雌性SD大鼠30只,用于建立脊髓损伤动物模型,随机分成单纯损伤组、细胞移植组、细胞移植+法舒地尔组,10只/组。法舒地尔为天津红日药业公司产品。方法:造模1周后,细胞移植组、细胞移植+法舒地尔组大鼠显露脊髓损伤区域,注入10μL骨髓间充质干细胞悬液。此外,细胞移植+法舒地尔组6h后开始腹腔注射10mg/kg法舒地尔,2次/d,连续用药1周。主要观察指标:斜板实验检测后肢运动功能,病理切片苏木精-伊红染色结果,HRP逆行神经示踪,Western-Blot法检测脊髓组织Phospho-ERM蛋白的表达。结果:①造模后8周,与单纯损伤组比较,细胞移植组及细胞移植+法舒地尔组大鼠斜板试验角度均显著增加(P<0.05,P<0.01),且后者增加幅度明显大于前者(P<0.05)。②单纯损伤组损伤处脊髓组织断裂,有明显空洞形成;细胞移植组、细胞移植+法舒地尔组可见少量神经轴索样结构,但细胞移植组的组织空洞大于细胞移植+法舒地尔组。③单纯损伤组至T8以上节段可见较少的HRP阳性颗粒标记的神经纤维;细胞移植组HRP阳性神经纤维数居中;细胞移植+法舒地尔组可见较多HRP阳性颗粒标记的神经纤维。④单纯损伤组、细胞移植组脊髓细胞Phospho-ERM蛋白的表达均明显高于细胞移植+法舒地尔组(P<0.05)。结论:骨髓间质干细胞移植对于脊髓损伤大鼠后肢运动功能的恢复有促进作用,联合应用法舒地尔可产生协同效果。
BACKGROUND： Research has been proved that Fasudil, a Rho kinase inhibitor, can effectively inhibit the onset of secondary spinal cord injury.
OBJECTIVE： To investigate the synergistic effect of bone marrow mesenchymal stem cell （BMSC） transplantation and Fasudil on motor functional recovery following spinal cord injury.
DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at Institute of Endocrinology, Tianjin Medical University from November 2008 to March 2009. MATERIALS： A 1-month-old SD rat was obtained to extract BMSCs. Another 30 healthy female SD rats were used to establish spinal cord injury models, and they were then randomly divided into single injury group, cell transplantation group, and cell transplantation ＋ Fasudil group, with 10 rats for each group. Fasudil was provided by Tianjin Hongri Pharmaceutical Co., Ltd.
METHODS： One week after modeling, spinal cord injury was exposed in the cell transplantation group and cell transplantation ＋ Fasudil group, and 10 μL BMSC suspension was inserted into the injured region. Otherwise, 6 hours later rats in the cell transplantation ＋ Fasudil group were treated with an intraperitoneal injection of 10 mg/kg Fasudil, twice a day for one successive week.
MAIN OUTCOME MEASURES： Hindlimb motor function was detected using inclined plane test. The Phosphorylated-ERM protein expression was detected by hematoxylin-eosin staining, pathology and horseradish peroxidase （HRP） nerve trace, and Western blot.
RESULTS： Eight weeks after modeling, degree of inclined plane was significantly increased in cell transplantation group and cell transplantation ＋ Fasudil group compared with single injury group （P 〈 0.05, P 〈 0.01）; while the increased value in the cell transplantation group was significantly greater than cell transplantation ＋ Fasudil group （P 〈 0.05）. Broken myeloid tissue and cavitation were observed in the single injury group; a few of neuraxis-like structures were observed in the cell transplantation group and cell transplantation ＋ Fasudil group, but the cavity in the cell transplantation group was larger than cell transplantation ＋ Fasudil group. HRP-positive nerve fibers were detected at T8 segment or even above in the single injury group and increased in cell transplantation group, in particular in cell transplantation ＋ Fasudil group. Phospho-ERM protein expression in the single injury group and cell transplantation group was significantly greater than cell transplantation ＋ Fasudil group （P 〈 0.05）.
CONCLUSION： BMSC transplantation can promote hindlimb motor functional recovery following spinal cord injury, while the combined application of cell transplantation and Fasudil may cause a synergistic effect.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第45期8901-8905,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
