NF-κB配体受体激动因子在脊髓损伤后骨髓间充质干细胞移植中的作用

目的探讨NF-κB配体受体激动因子(receptor activator of NF-kappa B ligand,RANKL)在脊髓损伤后人骨髓间充质干细胞(hBMSCs)移植中对神经功能恢复的影响。方法Wistar大鼠60只,体重250～350 g,雌雄不限,随机分为3组:(1)hBMSCs组,脊髓半横切后行hBMSCs移植;(2)RANKL hBMSCs组,脊髓半横切后hBMSCs和RANKL协同移植;(3)PBS组,脊髓半横切,移植物以PBS代替。移植后1,7,14,21,28 d采用行为学指标观察大鼠神经功能恢复情况,应用免疫组化和免疫荧光技术检测移植的BrdU标记hBMSCs在宿主脊髓内胶质纤维酸性蛋白(GFAP)和神经元特异性核蛋白(NeuN)的表达情况。结果大鼠脊髓半横切损伤后,hBMSCs组和RANKL hBMSCs组均较PBS对照组有明显的神经功能恢复;而移植7,14 d时,RANKL hBMSCs组动物较hBMSCs组神经功能恢复明显。RANKL hBMSCs组中hBMSCs-Ne- uN和hBMSCs-GFAP双标阳性率高于hBMSCs组。结论RANKL和hBMSCs协同移植可提高脊髓损伤后神经病学预后,提高hBMSCs在宿主体内向神经细胞的分化。

Effect of RANKL on intraspinal transplantation of bone marrow mesenchymal stem cells after spinal cord injury in rats

Objective To observe the effect of receptor activator of NF kappa B ligand (RANKL) on intraspinal transplantation of bone marrow mesenchymal stem cells (hBMSCs) after spinal cord injury in adult rats.Methods Sixty adult Wistar rats,weighing 250 ~ 350g,were used for the ex- periment.They were subjected to left spinal cord hemisection,then divided into three groups:( 1 ) hBM- SCs group,BrdU-labeled hMSCs were injected adjacent to the injury area;(2) RANKL hBMSCs group,BrdU -labeled hMSCs with RANKL were injected adjacently to the injury area;(3) PBS group, phosphate-buffered saline (PBS) was injected adjacently to the injury area.Functional outcome measure- ments using the modified Tarlov score were performed at 1,7,14,21 and 28 days after transplantation. After the animals were killed,spinal cord sections were examined using immunohistochemical and immu- nofluorescence staining methods to detect if BrdU - labeled adult hBMSCs express the astrocytic marker- GFAP (glial fibriliary acidic protein) and the neuron marker-NeuN (neuronal nuclei ),and compared be- tween the three groups.Results A statistically significant improvement in neurological function in rats that underwent hBMSCs and hBMSCs plus RANKL transplantation was found at each time point,com- pared with PBS injection rats.Meanwhile,a better neurological functional outcome was found in group hBMSCs RANKL than in group hMSCs at 7 and 14 days.More BrdU-NeuN and BrdU-GFAP immunore- active hBMSCs were determined in rats that underwent hBMSCs and RANKL transplantation than hBMSCs injection only.Conclusion Intraspinal hBMSCs transplantation along with RANKL improves neurologi- cal outcome after spinal cord injury,and facilitates hBMSCs differentiation into neural cells.