脊髓干细胞移植对脊髓损伤后CD-95和p75表达及神经细胞凋亡的影响

Effects of spinal cord-derived neural stem cell transplantation of the expression of CD-95 and p75 and apoptosis following spinal cord injury

目的探讨胚胎脊髓神经干细胞移植对大鼠脊髓损伤后CD-95和p75表达及神经细胞凋亡的影响。方法160只SD大鼠随机分为空白组,假手术组,脊髓损伤组,细胞移植组,分别在细胞移植后1、3 d、1、2、4周应用苏木素-伊红(HE)染色)、免疫组织化学、Tunel(凋亡测定)标记、电镜方法观察CD-95、p75和神经细胞凋亡的变化。结果移植后1、3 d、1、2、4周,脊髓损伤组和细胞移植组比较,CD95(P<0.05);p75,P<0.01。Tunel方法测定细胞凋亡结果,差异有统计学意义(P<0.05)。电镜观察可见,空白对照组和假手术组未见凋亡现象,细胞移植后1周,损伤对照组电镜下可见大量神经细胞凋亡小体。细胞移植组未见凋亡小体。结论脊髓源性神经干细胞移植后减少了CD-95和p75的表达,也减少了凋亡,说明移植促进了脊髓神经功能的恢复,也抑制了继发性脊髓损伤。

Objective To study the effects of spinal cord-derived neural stem cell transplantation on the expression of CD-95 and p75 and apoptosis following the spinal cord injury （SCI）. Methods One hundred and sixty S-D rats weighing 200-250 g were randomly divided into cell transplantation group, sham operation group, simple injury group and vacant group. After the spinal cord-derived neural stem cells were separated and cultured in vitro to the fifth generation, the cells were transplanted into transactionally the injured spinal cord. The rats in the simple injury group and sham operation group were injected with the same dose of saline as in the transplantation group. The animal were sacrificed at 1 st day ,3 rd day, 1 st week ,2nd week and 4th week after transplantation,and the expression of CD-95 and p75 and the apoptosis of neural stem cells were observed Results Immunohistochemical stains in the sham operation group and vacant group, the results were normal. Atl d after transplantation, CD-95 in the transplantation and simple injury group,（32.0±4.3）, （63.2± 1.2）;p75 （35.0±2.7）, （64.0 ＋ 1.3）. At 3d,CD-95, （48.0±2.3）, （74.0±1.5） ;p75, （40.0±2.1）, （81.0±3.2）. At 1 week,CD_95, （53.0± 1.4）, （110.0±3.2） ; p75, （47.0 ±2.5） , （94.0 ± 3.4）. At 2 weeks, CD_95, （30.0 ± 3.1）, （ 110.0 ± 3.2） ; p75, （32.0 ± 1.5） ,（62.0±2.3）. At 4 weeks,CD 95, （15.0± 1.8）, （21.0± 2.6） ;p75, （18.0± 3.1）, （35.0± 6.3 ）. Tunel, in the transplantation and simple injury group, at ld after transplantation, （ 16.0 ± 1.8） , （31.0 ±2.0） ;at 3d,（34.0±2.3） ,（71.0± 1.2） ;at 1 week,（42.0± 1.1） ,（93.0±3.1） ;at 2 weeks, （39.0 ±3.4） ,（81.0 ±2.1） ;at 4 weeks, （24.0 ±4.3） , （41.0 ±2.2）. Electron microscope： Apoptosis cells is obviously diminished in the transplantation group than the control group. Conclusions The cultured spinal cord derived cells repaired the injured spinal cord to some extent.