干细胞移植联合电针治疗对脊髓损伤修复的影响

Effect of treatment with stem cell transplantation combined with electric acupuncture on repair of spinal cord injury

目的研究人脐血干细胞移植联合电针刺激治疗脊髓损伤过程中对于类胰岛素样生长因子-1(IGF-1)表达的影响。方法将SD大鼠利用随机数字表法随机分成3组,所有大鼠均为脊髓横断损伤。脊髓损伤(SCI)组,干细胞移植(UCBSC)组,干细胞移植+电针刺激(UCBSC+EA)组,每组均为12只。UCBSC、UCBSC+EA组在损伤处注射脐血干细胞,SCI组以等量PBS缓冲液代替,UCBSC+EA组每天固定时间取损伤处上、下部位的"大椎""命门"进行电针刺激。各组均在损伤后的7天、14天和28天于损伤处取材,观察神经元细胞的形态、数量及IGF-1表达。结果常规HE组织学检查发现横断处灰质内神经元细胞变性、减少,并伴星形胶质细胞增生,细胞体肥大。免疫组化显示,损伤后IGF-1表达均上调,UCBSC、UCBSC+EA两组表达进一步上调,IGF-1的表达在前14天显著上调,14天后表达放缓。Western Blot结果显示,与单纯的人脐带血移植(UCBSC)相比较,UCBSC+EA更能促进IGF-1的表达,且有统计学意义(P<0.05)。RT-PCR电泳结果显示UCBSC+EA组,IGF-1mRNA的表达明显高于其他两组,有显著性差异(P<0.05)。结论 UCBSC+EA在大鼠脊髓损伤处可更有效分泌神经营养因子IGF-1,弥补脊髓损伤后维持神经元细胞生存所匮乏的营养因子,促进神经元细胞存活。

Objective To study the effect of the expression of spinal insulin-like growth factor-1 on treatment with human umbilical cord blood stem cell transplantation combined with electric acupuncture in spinal cord injury （SCI） rats. Methods All spinal cord injury SD rats were randomly divided into 3 groups , spinal cord injury groupl SCI）, umbilical cord blood stem cell group（UCBSC） and umbilical cord blood stem cell plus electric acupuncture group（UCBSCq-EA）（n： 12 ）. Umbilical cord blood stem cells were transplanted into spinal cord in the UCBSC group and the UCBSCq-EA group,while PBS was injected into the rats of the SCI group. After transplantation,rats in the UCBSCq-EA group were electro--acupunctured at a fixed time points every day. At the 7 d, 14 d and 28d,we observed the number of neurons,form and the expression of IGF-1. Results HE stai ning revealed that neuronal cells degenerated and reduced in the spinal cord matter ,while astrocytes had hyperplasia and hyper- trophy due to injury. The expression of IGF-1 was assayed by immunohistochemical staining in all groups ,especially UCBSC group and UCBSCq-EA group. The expression of IGF-1 increased significantly in 14 days, and slowdown after 14 days. West- ern Blot results showed that umbilical cord blood stem cell plus electric acupuncture promoted the expression of IGF-1 compared with simple human umbilical cord blood transplantation significantly（P〈0.05）. The expression of IGF-1 mRNA in the UCBSC q-EA group was significantly higher than other two groups （P〈0.05）. Conclusion In the UCBSCq-EA group,the rat spinal cord can secrete IGF-1 more effectively,make up the nutrition factor,and promote neuron cells survival.