CAJ | 학술논문

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6 h后开始腹腔注射10 mg/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.⑨单纯损伤组至T_8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.
배경:연구증실,Rho격매억제제법서지이가이유효조단계발성척수손상과정.목적:관찰법서지이대골수간충질간세포이식치료척수손상대서운동공능회복적영향.설계、시간급지점:수궤대조동물실험,우2008-11/2009-03재천진의과대학내분비연구소완성.재료:1월령SD대서1지,용우분리제비골수간충질간세포.건강자성SD대서30지,용우건립척수손상동물모형,수궤분성단순손상조、세포이식조、세포이식+법서지이조,10지/조.법서지이위천진홍일약업공사산품.방법:조모1주후,세포이식조,세포이식+법서지이조대서현로척수손상구역,주입10μL골수간충질간세포현액.차외,세포이식+법서지이조6 h후개시복강주사10 mg/kg법서지이,2차/d,련속용약1주.주요관찰지표:사판실험검측후지운동공능,병리절편소목정-이홍염색결과,HRP역행신경시종,Western-Blot법검측척수조직Phospho-ERM단백적표체.결과:①조모후8주,여단순손상조비교,세포이식조급세포이식+법서지이조대서사판시험각도균현저증가(P＜0.05,P＜0.01),차후자증가폭도명현대우전자(P＜0.05).②단순손상조손상처척수조직단렬,유명현공동형성;세포이식조、세포이식+법서지이조가견소량신경축색양결구,단세포이식조적조직공동대우세포이식+법서지이조.⑨단순손상조지T_8이상절단가견교소적HRP양성과립표기적신경섬유:세포이식조HRP양성신경섬유수거중;세포이식+법서지이조가견교다HRP양성과립표기적신경섬유.④단순손상조、세포이식조척수세포Phospho-ERM단백적표체균명현고우세포이식+법서지이조(P＜0.05).결론:골수간질간세포이식대우척수손상대서후지운동공능적회복유촉진작용,연합응용법서지이가산생협동효과.
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 masenchymal stem cell (BMSC) transplantion and Fasudil on motor functional recovery following spinal cord injury.DESIGN,TIME AND SETTING:A randomized controlled animal experimant 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 hernatoxylin-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-pesitive nerve fibers were detected at T_8 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.