自体骨髓干细胞动员移植与手术移植治疗脊髓损伤的实验研究  

Experimental study on autologous marrow stem cells mobilization and surgical transplantation for treatment of spinal cord injury

作　　者：李志营[1] 步星耀[1] 张圣旭[1] 刘猛[1] 程培训[1] 

机构地区：[1]河南省人民医院神经外科,郑州450003

出　　处：《中华临床医师杂志（电子版）》2010年第9期99-104,共6页Chinese Journal of Clinicians(Electronic Edition)

基　　金：河南省杰出人才计划项目(084200410011)

摘　　要：目的探讨自体骨髓干细胞(BMSC)动员移植与手术移植治疗脊髓损伤(SCI)的疗效和机制。方法选用SD大鼠90只,建模前注射溴脱氧尿嘧啶核苷(Brdu)50mg·kg-1·d-1×3d后抽取自体骨髓,体外分离自体BMSC;New York University(NYU)Impactor制作SCI模型,随机分为对照组、动员移植组、手术移植组各30只。动员移植组应用G-CSF皮下注射,20mg·kg-1·d-1×7d;手术移植组为损伤局部移植0.3ml(1×107个/ml)BMSC。采用BBB评分检测大鼠后肢的运动功能;SEP和MEP检测脊髓上、下行神经传导通路,判断SCI和恢复程度;脊髓组织病理和免疫组织化学检测Brdu、GFAP、BDNF、NSE和FactorⅧ抗原,TUNEL检测细胞凋亡。结果 BBB评分1周以后动员移植组和手术移植组分别较对照组比较差异有统计学意义(P<0.05),SEP、MEP潜伏期和波幅2周后动员移植组和手术移植组较对照组比较差异有统计学意义(P<0.05),组织病理学显示动员移植组和手术移植组较对照组有更少的空洞、坏死及GFAP阳性胶质瘢痕组织,较多的Brdu阳性细胞和NSE阳性细胞。BDNF表达和微血管计数均较对照组显著增多(P<0.05),动员移植组和手术移植组细胞凋亡数较对照组显著减少(P<0.05)。结论自体BMSCs动员移植和手术移植两种方法均能使BMSCs向脊髓损伤组织区聚集、增殖、分化,分泌神经营养因子、促进血管生成、减少细胞凋亡,促进神经再生与TBI修复,改善大鼠脊髓功能,促进损伤后的脊髓功能的恢复;二者对比,前者更为方便、无创,实用性强,更有可能抓住有限的治疗时机,因而应用前景可能更好。Objective To observe the effect of treating spinal cord injury（SCI） in rats by mobilization and surgical transplantation of autologous bone marrow stem cells （BMSC）.Methods Ninety healthy SD rats,half male and half female,were randomly divided into control group,mobilized transplanted group and surgically transplantated group.Rats in mobilized transplanted group were subcutaneously injected with granulocyte-colony stimulating factor （granulocyte-colony stimulating factor,G-CSF） for 7 days.After 3 days injection with Bromodeoxyuridine,BMSCs were isolated from healthy rats.All the rats were underwent spinal cord injury by NYU impactor.In surgical transplantation group,a total of 0.3 ml（1 × 107 /ml） BMSC were transplanted into injured area of spinal cord.All the rats were injected with 50 mg/kg Bromodeoxyuridine （50 mg/kg） in the following 10 days.The BBB locomotor score were used to evaluate functional recovery in rat hindlima.MEP and SEP test were performed to evaluate recovery of injured spinal cord.The extent of injury,bleeding,cellular necrosis and fibrosis were investigated by HE dyeing.Immunohistochemistry detects BrdU,GFAP,BDNF,NSE and Factor Ⅷ antigen.Terminal deoxynucleotidyl Transferase Biotin-dUTP Nick End Labeling （TUNEL） detects apoptosis.Results BBB score was higher in mobilization group and surgical transplantation group than the control group 1 weeks after injury,the statistical difference was significant（P 〈 0.05）,statistical difference showed in latency period and wave amplitude in SEP＆MEP after 2 weeks,the BDNF positive cells of mobilized transplanted group and surgically transplantated group were significantly more than that of control group （P 〈 0.05）,while the apoptotic cells are less than that of the control group.Conclusions Autologous BMSC mobilization transplants and surgery transplant could significantly reduce the degree of SCI,and promote the recovery of the spinal cord function after injury for treating SCI.Mobilization of autologous BMSC are mo

关 键 词：脊髓损伤 骨髓祖代细胞 细胞移植 模型 动物 

分 类 号：R651.2[医药卫生—外科学]