CAJ | 학술논문

目的探讨间充质干细胞(MSCS)移植对大鼠脊髓损伤后脑源性神经营养因子(BDNF)表达的影响.方法选取SD大鼠4只作骨髓间充质细胞的分离与培养,提取MSCS;60只大鼠制做脊髓横断损伤模型,细胞移植组24只,PBS(磷酸盐缓冲液)液组24只,空白对照组12只.于脊髓损伤后第7天,无菌条件下,细胞移植组以微量注射器缓慢注入含MSCS的培养液5μl,PBS组注入缓冲液体5 μl,对照组未加任何干预因素.分别于术后7、14、28 d麻醉下行心脏灌流固定取T10节段脊髓,细胞移植组与磷酸盐缓冲组取出损伤节段的脊髓(8只/时点),空白对照组于同一节段取出相应脊髓(4只/时点).应用免疫组化法观察MSCS移植后大鼠脊髓损伤区BDNF的表达变化.结果细胞接种24 h后贴壁生长,72 h细胞增殖,有细胞团形成,在传代过程中,4代以前的细胞倍增时间约为4～6 d,至10代以后细胞增殖能力有所减弱,胞体变得扁平,若加入碱性成纤维细胞生长因子(bFGF),则可维持基增殖能力和形态.脑源性神经营养因子在正常大鼠脊髓组织中有一定表达,间充质干细胞移植术后第7、14及28天,细胞移植组脑源性神经营养因子均高水平表达,与缓冲液组相比较差异有统计学意义(14 d:0.31±0.03 vs 0.25±0.04,P＜0.01).结论间充质干细胞在移植后通过上调脑源性神经营养因子的表达从而促进轴突的再生可能是治疗脊髓损伤的重要机制.
목적 탐토간충질간세포(MSCS)이식대대서척수손상후뇌원성신경영양인자(BDNF)표체적영향.방법 선취SD대서4지작골수간충질세포적분리여배양,제취MSCS;60지대서제주척수횡단손상모형,세포이식조24지,PBS(린산염완충액)액조24지,공백대조조12지.우척수손상후제7천,무균조건하,세포이식조이미량주사기완만주입함MSCS적배양액5μl,PBS조주입완충액체5 μl,대조조미가임하간예인소.분별우술후7、14、28 d마취하행심장관류고정취T10절단척수,세포이식조여린산염완충조취출손상절단적척수(8지/시점),공백대조조우동일절단취출상응척수(4지/시점).응용면역조화법관찰MSCS이식후대서척수손상구BDNF적표체변화.결과 세포접충24 h후첩벽생장,72 h세포증식,유세포단형성,재전대과정중,4대이전적세포배증시간약위4～6 d,지10대이후세포증식능력유소감약,포체변득편평,약가입감성성섬유세포생장인자(bFGF),칙가유지기증식능력화형태.뇌원성신경영양인자재정상대서척수조직중유일정표체,간충질간세포이식술후제7、14급28천,세포이식조뇌원성신경영양인자균고수평표체,여완충액조상비교차이유통계학의의(14 d:0.31±0.03 vs 0.25±0.04,P＜0.01).결론 간충질간세포재이식후통과상조뇌원성신경영양인자적표체종이촉진축돌적재생가능시치료척수손상적중요궤제.
Objective To observe the effects of mesenchymal stem cells (MSCS) transplantation on the brain-derived neurotrophic factors (BDNF) after the spinal cord injury (SCI) of rats, and investigate the mechanism of repairing the SCI by MSCS transplantation.Methods Mesenchymal stem cells were cultured from the thighbone of adult SD rats and identified by immunoctochemistry.Seven days after the operation of spinal cord injury, the mesenchymal stem cells were transplanted into the injured spinal cord site.Sixty adult SD rats were random divided into three groups: Spinal cord injury cured with transplants of mesenchymal stem cells to the injured spinal cord site ( group A), spinal cord injury received PBS solution( group B) and control group ( group C).The expression of brain - derived neurotrophic factors of the lesion and neighbor areas were examined by immunohistochemistry, The mechanism of repairing the lesion and neighbor areas were examined by immunohistochemistry, and the mechanisms of repairing the spinal cord injury after mesenchymal stem cells transplantation were investigated.Results Mesenchymal stem cells began to grow after 24 hours of inoculation and proliferate 72 hours later.The proliferate the proliferous cyclewas 4 ～6 days, it declined on the 10th era.Mesenchymal stem cells turned to flat, and the mesenchymal stem cells could maintain the ability of proliferation if bFGF was added.Compared with group B, transplantation of mesenchymal stem cells enhanced more expression of brain-derived neurotrophic factors in group A ( 14 d :0.31 ± 0.03 vs 0.25 ± 0.04, P ＜ 0.01 ).Conclusion Mesenchymal stem cells in transplantation group showed a continued high level of brain-derived neurotrophic factors, which may be one of the mechanisms of repairing the spinal cord injury by mesenchymal stem cells transplantation.