CAJ | 학술논문

背景:干细胞治疗神经组织损伤的关键在于移植具有再生能力的干细胞,通过多种作用机制,重建中枢神经系统的结构和功能.目的:立体定向移植骨髓间充质干细胞观察其对大鼠脊髓损伤修复的影响并探讨其机制.方法:密度梯度离心法结合贴壁筛选法分离骨髓间充质干细胞,于细胞移植前掺入10 mg/L的BrdU进行标记.将成年雌性Wistar大鼠以动脉瘤夹闭法制备大鼠脊髓损伤模型后随机分为对照组、生理盐水组与移植组.移植组大鼠致伤后第7天,通过立体定向途径移植骨髓间充质干细胞到脊髓损伤中心,生理盐水组给予等量生理盐水,对照组大鼠不做处理.于大鼠脊髓损伤前及损伤后第7,14,30,60,90天进行BBB评分;损伤后第90天处死大鼠,观察其脊髓组织中有无BrdU阳性细胞、Brdu+神经元特异性烯醇化酶、Brdu+胶质纤维酸性蛋白、Brdu+碱性成纤维细胞生长因子、Brdu+脑源性神经生长因子免疫组织化学双染阳性细胞及单染阳性细胞.结果与结论:①骨髓间充质干细胞移植组大鼠BBB后肢功能评分恢复优于对照组(P＜0.05):生理盐水组BBB评分在损伤后30 d内恢复速度慢于对照组(P＜0.05),至第90天与对照组比较差异无显著性意义(P＞0.05).②移植组大鼠脊髓内在损伤中心及头、尾端距离脊髓损伤中心1 cm处均可见BrdU染色阳性细胞及免疫组织化学双染阳性细胞.移植组神经元特异性烯醇化酶、胶质纤维酸性蛋白、碱性成纤维细胞生长因子、脑源性神经生长因子单染阳性细胞数明显高于对照组和生理盐水组(P＜0.05).结果提示,骨髓间充质干细胞移植可以促进脊髓损伤大鼠的神经功能的恢复,其机制可能与移植细胞分化为神经元样和神经胶质细胞样细胞,并分泌或促进宿主分泌神经营养因子有关.
배경:간세포치료신경조직손상적관건재우이식구유재생능력적간세포,통과다충작용궤제,중건중추신경계통적결구화공능.목적:입체정향이식골수간충질간세포관찰기대대서척수손상수복적영향병탐토기궤제.방법:밀도제도리심법결합첩벽사선법분리골수간충질간세포,우세포이식전참입10 mg/L적BrdU진행표기.장성년자성Wistar대서이동맥류협폐법제비대서척수손상모형후수궤분위대조조、생리염수조여이식조.이식조대서치상후제7천,통과입체정향도경이식골수간충질간세포도척수손상중심,생리염수조급여등량생리염수,대조조대서불주처리.우대서척수손상전급손상후제7,14,30,60,90천진행BBB평분;손상후제90천처사대서,관찰기척수조직중유무BrdU양성세포、Brdu+신경원특이성희순화매、Brdu+효질섬유산성단백、Brdu+감성성섬유세포생장인자、Brdu+뇌원성신경생장인자면역조직화학쌍염양성세포급단염양성세포.결과여결론:①골수간충질간세포이식조대서BBB후지공능평분회복우우대조조(P＜0.05):생리염수조BBB평분재손상후30 d내회복속도만우대조조(P＜0.05),지제90천여대조조비교차이무현저성의의(P＞0.05).②이식조대서척수내재손상중심급두、미단거리척수손상중심1 cm처균가견BrdU염색양성세포급면역조직화학쌍염양성세포.이식조신경원특이성희순화매、효질섬유산성단백、감성성섬유세포생장인자、뇌원성신경생장인자단염양성세포수명현고우대조조화생리염수조(P＜0.05).결과제시,골수간충질간세포이식가이촉진척수손상대서적신경공능적회복,기궤제가능여이식세포분화위신경원양화신경효질세포양세포,병분비혹촉진숙주분비신경영양인자유관.
BACKGROUND:The key of stem cells for treating nervous tissue injury is the transplantation of stem coils that have regeneration capacity.The structure and function of central nervous system were re-established by multiple action mechanisms.OBJECTIVE:To explore the effects and mechanisms of bone marrow mesenchymal stem cells(BMSCs)locally transplanted into rats with spinal cord injury on neurological recovery.METHODS:BMSCs were separated with density gradient centrifugation and cell attachment.10 mg/L BrdU was used for labeling before cell transplantation.Adult female Wistar rats were used to establish spinal cord injury models using an aneurysm clip,and they were then randomly divided into control group,saline group and transplantation group.In the transplantation group,BMSCs were transplanted into the damaged spinal cord by stereotaxis at day 7 following damage.In the saline group,an equal volume of saline was utilized.In the control group,the rats were left intact.Basso,Beattie and Bresnahan(BBB)locomotor rating scale was used before and at 7,14,30,60 and 90 days following damage.Rats were sacrificed at day 90.BrdU-positive cells,Brdu+neuron specific enolase,Brdu+glial fibrillary acidic protein(GFAP),Brdu+basic fibroblast growth factor(bFGF),and Brdu+brain-derived nerve growth factor(BDNF)immunohistochemistry double-staining cells and simple staining positive calls were observed.RESULTS AND CONCLUSION:The recovery of BBB function score was better in the transplantation group than in the control group(P＜0.05).The recovery speed of BBB function score was slower in the saline group than in the control group at 30 days following damage(P＜0.05).No significant difference was determined at day 90 compared with the control group(P＞0.05).BrdU-positive cells and double-staining cells of immunohistochemistry could be found at the center of damage site and 1 cm from caudal end to damaged site in rats of the transplantation group.The number of NSE,GFAP,bFGF and BDNF simple staining cells was significantly greater in the transplantation group than in the control and saline groups(P＜0.05).Results indicated that BMSC transplantation can improve the recovery of nervous function of rats with spinal cord injury.Its mechanism may be correlated with the differentiation of transplanted cells into neuron-like and glial cell-like cells,secretion or promoting secretion of neurotrophic factors in host.