过表达胶质细胞源性神经营养因子的脂肪源性间充质干细胞对大鼠电损伤坐骨神经的作用  

Effects of adipose-derived mesenchymal stem cells over-expressing glial cell line-derived neurotrophic factor on electrically injured sciatic nerve of rats

作　　者：杨晨[1] 胡大海[1] 郑朝[1] 白晓智[1] 王耀军[1] 汤朝武[1] 

机构地区：[1]第四军医大学西京医院全军烧伤中心、烧伤与皮肤外科,西安710032

出　　处：《中华烧伤杂志》2015年第3期199-204,共6页Chinese Journal of Burns

基　　金：国家自然科学基金（81471879）

摘　　要：目的观察持续过表达胶质细胞源性神经营养因子（GDNF）的脂肪源性间充质干细胞（ADSC）对大鼠坐骨神经电刺激损伤后肢体运功功能恢复和神经修复的影响。方法取5只SD大鼠制备过表达GDNF的ADSC。取150只大鼠按照随机数字表法分为正常对照组、GDNF-ADSC组、ADSC组、GDNF组、生理盐水组5组，每组30只。正常对照组不作处理，进行常规饲养；GDNF-ADSC组、ADSC组、GDNF组、生理盐水组大鼠在右后肢大腿造成坐骨神经电损伤，伤后即刻向损伤神经表面分别局部注射100μL过表达GDNF的ADSC悬液（细胞浓度1×10 7个／mL）、ADSC细胞悬液（细胞浓度1×10 7个／mL）、GDNF溶液（100mg／L）、生理盐水。伤后第1—8周，每组取6只大鼠进行后肢步幅测量，并观察其伤后第8周的坐骨神经形态；伤后第4周，采用蛋白质印迹法检测各组剩余大鼠坐骨神经的GDNF蛋白表达量。对数据行单因素方差分析、重复测量方差分析、SNK检验。结果GDNF-ADSC组、ADSC组、GDNF组、生理盐水组大鼠在伤后各时相点后肢步幅均明显短于正常对照组（P值均小于0．05）；GDNF—ADSC组大鼠在伤后第3—8周、ADSC组大鼠在伤后第3、5、7周、GDNF组大鼠在伤后第3、5、7、8周后肢步幅均明显长于生理盐水组（P值均小于0．05）；伤后第4—8周，GDNF-ADSC组大鼠后肢步幅分别为（10．83±0．97）、（13．25±1．40）、（12．86±1．42）、（14．06±1．50）、（15．09±1．17）cm，明显长于ADSC组的（8．90±0．82）、（9．03±0．57）、（9．27±0．36）、（9．86±0．36）、（9．52±0．58）em。和GDNF组的（8．87±0．69）、（8．51±1．18）、（9．34±0．87）、（9．76±0．67）、（9．50±1．22）cm（P值均小于0．05）。伤后第8周，与正常对照组比较，生理盐水组大鼠坐骨神经有髓神经纤维数明显减少，GDNF—ADSC组、ADSC组、GDNF组有髓神经纤维数明显增Objective To observe the effects of adipose-derived mesenchymal stem cells （ADSCs） with continous over-expression of glial cell line-derived neurotrophic factor （GDNF） on the motor function recovery and nerve regeneration of sciatic nerve of rats after electrical injury. Methods Five SD rats were collected to prepare ADSCs with over-expression of GDNF. One hundred and fifty SD rats were divided into normal control group （ N）, GDNF-ADSCs group （ GA）, ADSCs group （ A）, GDNF group （ G）, and physi- ological saline group （P） according to the random number table, with 30 rats in each group. Rats in group N were routinely fed without treatment, and rats in the other 4 groups were inflicted with electrical injury on sciatic nerve of thigh of the right hind leg. Rats in groups GA, A, G, and P were respectively injected with 100 μL suspension of ADSCs with over-expression of GDNF （ 1 x 107 cells per mL） , 100 μL ADSCs suspen- sion （ 1 x 107 cells per mL） , 100 μL GDNF solution （ 100 mg/L） , and 100 μL physiological saline to the surface of the injured nerves immediately after injury. Six rats of each group were collected for measuring hind limb stride from post injury week （PIW） 1 to 8, and morphology of the sciatic nerves was observed in PIW 8. In PIW 4, the protein expression of GDNF of sciatic nerves of the rest rats in each group was deter- mined with Western blotting. Data were processed with one-way analysis of variance, analysis of variance of repeated measurement, and SNK test. Results Compared with that of group N, the hind limb stride val- ues in groups GA, A, G, and P were significantly lower at each time point （ with P values below 0.05 ）. Compared with those of group P, the hind limb stride values in group GA from PIW 3 to 8, in group A in PIW 3, 5, and 7, and in group G in PIW 3, 5, 7, and 8 were significantly longer （ with P values below 0.05）. The hind limb stride values in group GA from PIW 4 to 8 were respectively （10. 83 ± 0. 97） , （13.25±1

关 键 词：烧伤 电 胶质细胞源性神经营养因子 干细胞 坐骨神经 慢病毒转染 

分 类 号：R647[医药卫生—外科学]