经脑源性神经营养因子基因修饰的脂肪间充质干细胞对大鼠急性脊髓损伤的影响  

Effect of brain-derived neurotrophic factor.genetically modified adipose derived mesenchymai stem cells on acute spinal cord injury in rats

作　　者：熊敏[1] 曾云[1] 刘志刚[1] 余化龙[1] 陈森[1] 何宏生[1] 

机构地区：[1]湖北医药学院附属东风医院骨科,十堰442000

出　　处：《中华实验外科杂志》2013年第5期1032-1034,共3页Chinese Journal of Experimental Surgery

基　　金：基金项目：湖北省教育厅资助项目（B20112106）

摘　　要：目的探讨慢病毒转染脑源性神经营养因子（BDNF）的脂肪间充质干细胞（ADSCs）脊髓移植的应用价值。方法体外分离培养大鼠ADSCs，用高滴度慢病毒将GDNF转染入ADSCs。75只SD大鼠随机分为3组，分别用BDNF基因修饰的ADSCs（A组）、ADSCs（B组）、生理盐水（C组）注射治疗。观察Basso-Beattie—Bresnahan（BBB）评分、免疫荧光染色及脑源性神经营养因子（BDNF）、神经生长因子（NGF）、胶质纤维酸性蛋白（GFAP）的表达。结果术后3、4周，A组运动功能评分为（15．02±1．25）、（17．43±1．05）分，明显高于B组（10．35±2．25）、（13．44±1．83）分、C组（5．88±2．46）、（6．94±3．22）分。免疫荧光显示移植脂肪干细胞脊髓组织内，均不同程度的可见分布有溴脱氧尿嘧啶核苷（BrdU）阳性标记细胞。术后3周，A组BDNF、NGF、GFAP表达分别为（38．70±2．75）、（35．62±2．74）、（13．17±3．09）个／HP，与B组（28．80±2．05）、（24．99±3．58）、（21．83±5．99）个／HP、C组（24．60±2．44）、（20．33±2．25）、（27．10±6．47）个／HP比较差异有统计学意义（P〈0．05）。结论ADSCs移植能促进损伤局部脊髓的BDNF、NGF的表达增加，并抑制GFAP的表达。BDNF基因修饰的ADSCs移植能更加强这一趋势。这可能是促进神经结构及神经功能恢复的因素之一。Objective To-investigate the applied value of brain-derived neurotrophic factor （BDNF） -transfected adipose-derived stem cells （ADSCs） by lentivims in spinal transplantation. Methods Rat ADSCs were isolated and cultured in vitro under sterile conditions. Then they were transfected with GDNF by high titer lentivirus. Thirty-six SD rats were randomly divided into 3 groups, respectively treated with ADSCs transfected with GDNF by lentivirus （ A）, ADSCs （ B）, normal saline （C）. Basso-Beattie- Bresnahan （BBB） score, immunofluorescence stain, and the expression levels of BDNF, nerve growth factor （NGF） and glial fibrillary acidic protein （GFAP） were observed. Results On the postoperative week （POW） 3 and 4, the BBB scores in group A were （ 15.02 ± 1.25） and （ 17. 43 ± 1.05） points, obviously higher than in group B （10.35 ±2.25） and （13.44 ±1.83） points and group C （5.88±2.46） and （ 6. 94 ± 3.22） points. The immunofluoreseenee stain revealed that the transplanted ADSCs migrated to the injury site. On POW 3, the BDNF, NGF and GFAP expression levels in group A were （38.70 ± 2. 75）, （35.62 ± 2. 74 ） and （ 13.17 ± 3.09 ） eells/HP, significantly different from those in group B （ 28.80 ± 2.05）, （24. 99 ± 3.58 ） and （21.83 ± 5.99） eells/HP and group C （24. 60 ± 2. 44）, （ 20. 33 ± 2. 25 ） and （27. 10 ± 6.47） cells/HP. Conclusion After transplantation of ADSCs in rats with spinal cord injury, the expression of BDNF and NGF was increased, and that of GFAP was suppressed, which was promoted by the modification of BDNF. Maybe it is one of the mechanisms of promoting restoration of injured spinal cord and improving motor function.

关 键 词：脑源性神经营养因子 脂肪间充质干细胞 急性脊髓损伤 

分 类 号：R651[医药卫生—外科学]