中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2012年
12期
1204-1208
,共5页
徐玉生%王培松%马玉斐%金伟林%李星辰%贾延劫%苗金红
徐玉生%王培鬆%馬玉斐%金偉林%李星辰%賈延劫%苗金紅
서옥생%왕배송%마옥비%금위림%리성신%가연겁%묘금홍
间充质干细胞%脊髓损伤%慢病毒载体
間充質榦細胞%脊髓損傷%慢病毒載體
간충질간세포%척수손상%만병독재체
Marrow mesenchymal stem cell%Spinal cord injury%Lentiviral vector
目的 通过观察miRNA-9在骨髓间充质干细胞(MSCs)分化为神经元过程中的作用,探讨基因修饰在脊髓损伤治疗中的作用.方法 分离培养大鼠MSCs并构建miRNA-9-1慢病毒载体.成功建立84只大鼠急性脊髓损伤模型,并按照随机数字表法分为对照组、MSCs组及miRNA组,每组28只.脊髓损伤后1周,对MSCs组大鼠进行MSCs移植,对miRNA组大鼠移植miRNA-9-1慢病毒载体感染的MSCs,对照组大鼠仅在损伤部位注射等量的生理盐水.选择不同时间点对大鼠后肢进行Basso Beattie Bresnahan(BBB)评分,并行神经丝蛋白200(NF-200)和胶质纤维酸性蛋白(GFAP)免疫组织化学染色,对各组阳性表达面积百分比进行比较.结果 细胞移植4周后,各组大鼠BBB评分差异有统计学意义,其中miRNA组评分较MSCs组及对照组明显提高,差异有统计学意义(P＜0.05).免疫组织化学染色示miRNA组的NF-200阳性面积较MSCs组及对照组明显增大,GFAP阳性面积较MSCs组及对照组明显减小,各组间的差异具有统计学意义(P＜0.05).结论 miRNA-9在MSCs横向分化为神经元中起重要调控作用,并通过促进轴突再生,减少脊髓损伤部位反应性胶质细胞的数量等机制促进脊髓损伤后的功能修复.
目的 通過觀察miRNA-9在骨髓間充質榦細胞(MSCs)分化為神經元過程中的作用,探討基因脩飾在脊髓損傷治療中的作用.方法 分離培養大鼠MSCs併構建miRNA-9-1慢病毒載體.成功建立84隻大鼠急性脊髓損傷模型,併按照隨機數字錶法分為對照組、MSCs組及miRNA組,每組28隻.脊髓損傷後1週,對MSCs組大鼠進行MSCs移植,對miRNA組大鼠移植miRNA-9-1慢病毒載體感染的MSCs,對照組大鼠僅在損傷部位註射等量的生理鹽水.選擇不同時間點對大鼠後肢進行Basso Beattie Bresnahan(BBB)評分,併行神經絲蛋白200(NF-200)和膠質纖維痠性蛋白(GFAP)免疫組織化學染色,對各組暘性錶達麵積百分比進行比較.結果 細胞移植4週後,各組大鼠BBB評分差異有統計學意義,其中miRNA組評分較MSCs組及對照組明顯提高,差異有統計學意義(P＜0.05).免疫組織化學染色示miRNA組的NF-200暘性麵積較MSCs組及對照組明顯增大,GFAP暘性麵積較MSCs組及對照組明顯減小,各組間的差異具有統計學意義(P＜0.05).結論 miRNA-9在MSCs橫嚮分化為神經元中起重要調控作用,併通過促進軸突再生,減少脊髓損傷部位反應性膠質細胞的數量等機製促進脊髓損傷後的功能脩複.
목적 통과관찰miRNA-9재골수간충질간세포(MSCs)분화위신경원과정중적작용,탐토기인수식재척수손상치료중적작용.방법 분리배양대서MSCs병구건miRNA-9-1만병독재체.성공건립84지대서급성척수손상모형,병안조수궤수자표법분위대조조、MSCs조급miRNA조,매조28지.척수손상후1주,대MSCs조대서진행MSCs이식,대miRNA조대서이식miRNA-9-1만병독재체감염적MSCs,대조조대서부재손상부위주사등량적생리염수.선택불동시간점대대서후지진행Basso Beattie Bresnahan(BBB)평분,병행신경사단백200(NF-200)화효질섬유산성단백(GFAP)면역조직화학염색,대각조양성표체면적백분비진행비교.결과 세포이식4주후,각조대서BBB평분차이유통계학의의,기중miRNA조평분교MSCs조급대조조명현제고,차이유통계학의의(P＜0.05).면역조직화학염색시miRNA조적NF-200양성면적교MSCs조급대조조명현증대,GFAP양성면적교MSCs조급대조조명현감소,각조간적차이구유통계학의의(P＜0.05).결론 miRNA-9재MSCs횡향분화위신경원중기중요조공작용,병통과촉진축돌재생,감소척수손상부위반응성효질세포적수량등궤제촉진척수손상후적공능수복.
Objective To assess the role of microRNA-9 in bone marrow mesenchymal stem cells differentiating into neurons and research the role of gene modification in spinal cord injury treatment.Methods Adherent culture was used to isolate and culture rat bone marrow mesenchymal stem cells (MSCs); microRNA-9-1 lentiviral vector was constructed.Acute spinal cord injury (SCI) models were established in 84 adult SD rats at T10/11 level according to the improved Allen's method; then,they were randomly divided into control group,MSCs group and miRNA group (n=28).One week after SCI,the rats of MSCs group were treated with MSCs implantation,and the rats of miRNA group were treated with MSCs-transfected microRNA-9-1 lentiviral vector; while rats of the control group only received the same amount of physical saline in the same region.The neurological functions were evaluated by using Basso-Beattie-Bresnahan (BBB) scale 1 and 3 days,and 1,2,4,6,8 and 12 weeks after SCI.The immunoreactivity of neuro filament 200 (NF-200) and glial fibrillary acidic protein (GFAP) was measured,and the percentage of positive response area was assayed and compared between groups.Results Four weeks after cell transplantation,statistical difference of BBB scale scores was noted between each two groups; the scores of miRNA group were significantly higher than those of the MSCs group and control group (P＜0.05).The immunohistochemistry staining indicated that the expression of NF-200 was significantly more intense and the expression of GFAP was significantly weaker in miRNA group than those of the other two groups (P＜0.05).Conclusion MicroRNA-9 may play an important role in bone marrow mesenchymal stem cells differentiating into neurons,and possess effects on repairing injured spinal cord and promoting functional recovery through promoting axonal regeneration and reducing the number of reactive glial cells in the spinal cord injury site.
