人脐带沃顿胶干细胞对急性大鼠脊髓损伤的作用及脊髓组织中TNF-α和BDNF表达的影响
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摘要
目的:探讨人脐带沃顿胶干细胞(WJCs)移植对急性脊髓损伤大鼠的治疗作用及其可能机制。方法:81只大鼠分为假手术组、模型组和WJCs移植组,每组27只。每组取6只,采用BBB评分法评估脊髓损伤后1、3、7、14、21和28 d各组大鼠的运动功能,采用透射电镜检测损伤局部超微结构并检测损伤后28 d脑源性神经营养因子(BDNF)的表达,其余大鼠采用实时荧光定量PCR和Western blot检测损伤后0、3、6、12、24、72、168 h损伤脊髓组织中肿瘤坏死因子(TNF-α)的表达。结果:脊髓损伤后模型组与WJCs移植组大鼠的运动功能均有不同程度的恢复,其中WJCs移植组较模型组恢复更明显。与模型组相比,WJCs移植组损伤区脊髓组织结构相对完整。模型组和WJCs移植组损伤脊髓组织中TNF-αmRNA和蛋白以及BDNF蛋白的表达较假手术组增加(P<0.05);与模型组相比,WJCs移植组中TNF-α的表达降低,而BDNF的表达增加(P<0.05)。结论:WJCs移植可改变脊髓损伤区的微环境,抑制损伤脊髓组织中TNF-α的表达,同时增加BDNF的表达,促进大鼠神经功能恢复,减少脊髓继发性损伤。
Aim: To explore the effect of Wharton's jelly cells( WJCs) transplant on the expressions of tumor necrosis factor alpha( TNF-α) and brain-derived neurotrophic factor( BDNF) in rats with acute spinal cord injury( SCI),and to investigate the function of WJCs in the inflammatory response and nerve repair after acute SCI. Methods: A total of 81 rats were allocated into sham operation group,model group and WJCs group( 27 rats in each group). In the WJCs group,200μL WJCs cell suspension( about 1 × 106cells) was injected by femoral vein at 10 min after the injury. Neurologic function was accessed by BBB score at 1 d,3 d,7 d,14 d,21 d and 28 d respectively after transplantation; the ultrastructures of injured spinal cord were detected by transmission electron microscopy; the mRNA and protein expressions of BDNF in the spinal cord tissue were detected by real-time quantitative PCR and Western blot,respectively at 28 d,and those of TNF-αlevel were detected at 0 h,3 h,6 h,12 h,24 h,72 h,168 h after transplantation. Results: Neural function was recovered to some extent both in WJCs group and model group,and the former was much better. Compared with model group,the structure of SCI area of WJCs group was relatively intact. Compared with the sham operation group,the expressions of BDNF protein and TNF-α mRNA and protein were increased significantly both in model group and WJCs group( P < 0. 05);compared with model group,the expressions of TNF-α mRNA and protein decreased( P < 0. 05),while that of BDNF increased in the WJCs group( P < 0. 05). Conclusion: WJCs transplantation can change the microenvironment of the injured spinal cord by inhibiting the expression of TNF-α while promoting the expression of BDNF in the lesion,therefore reducing secondary spinal cord injury,which may be a potential mechanism underlying the recovery of motor function of injured rats.
Aim: To explore the effect of Wharton's jelly cells( WJCs) transplant on the expressions of tumor necrosis factor alpha( TNF-α) and brain-derived neurotrophic factor( BDNF) in rats with acute spinal cord injury( SCI),and to investigate the function of WJCs in the inflammatory response and nerve repair after acute SCI. Methods: A total of 81 rats were allocated into sham operation group,model group and WJCs group( 27 rats in each group). In the WJCs group,200μL WJCs cell suspension( about 1 × 106cells) was injected by femoral vein at 10 min after the injury. Neurologic function was accessed by BBB score at 1 d,3 d,7 d,14 d,21 d and 28 d respectively after transplantation; the ultrastructures of injured spinal cord were detected by transmission electron microscopy; the mRNA and protein expressions of BDNF in the spinal cord tissue were detected by real-time quantitative PCR and Western blot,respectively at 28 d,and those of TNF-αlevel were detected at 0 h,3 h,6 h,12 h,24 h,72 h,168 h after transplantation. Results: Neural function was recovered to some extent both in WJCs group and model group,and the former was much better. Compared with model group,the structure of SCI area of WJCs group was relatively intact. Compared with the sham operation group,the expressions of BDNF protein and TNF-α mRNA and protein were increased significantly both in model group and WJCs group( P < 0. 05);compared with model group,the expressions of TNF-α mRNA and protein decreased( P < 0. 05),while that of BDNF increased in the WJCs group( P < 0. 05). Conclusion: WJCs transplantation can change the microenvironment of the injured spinal cord by inhibiting the expression of TNF-α while promoting the expression of BDNF in the lesion,therefore reducing secondary spinal cord injury,which may be a potential mechanism underlying the recovery of motor function of injured rats.
引文
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[13]张群,王竞秋,宋焱峰,等.大鼠急性脊髓损伤后MCP-1及BDNF表达变化的实验研究[J].甘肃医药,2013,32(3):184
[14]徐玉生,张松,金伟林,等.褪黑素对大鼠脊髓损伤后肿瘤坏死因子-α表达的影响[J].郑州大学学报:医学版,2014,49(1):41
[2]Zhang ZJ,Cao DL,Zhang X,et al.Chemokine contribution to neuropathic pain:respective induction of CXCL1 and CXCR2 in spinal cord astrocytes and neurons[J].Pain,2013,154(10):2185
[3]Hermann GE,Rogers RC,Bresnahan JC,et al.Tumor necrosis factor-alpha induces c FOS and strongly potentiates glutamate-mediated cell death in the rat spinal cord[J].Neurobiol Dis,2001,8(4):590
[4]Andrade P,Visser-Vandewalle V,Hoffmann C,et al.Role of TNF-alpha during central sensitization in preclinical studies[J].Neurol Sci,2011,32(5):757
[5]Hiebert GW,Khodarahmi K,Mcgraw J,et al.Brain-derived neurotrophic factor applied to the motor cortex promotes sprouting of corticospinal fibers but not regeneration into a peripheral nerve transplant[J].J Neurosci Res,2002,69(2):160
[6]邓许勇,周荣平,鲁凯伍,等.氯化锂联合脐血干细胞移植修复大鼠脊髓损伤[J].南方医科大学学报,2010,30(11):2436
[7]张文进,黄华,关方霞,等.人脐带间充质干细胞治疗脊髓损伤及其对内源性细胞增殖的影响[J].实用医学杂志,2013,29(6):880
[8]Basso DM,Beattie MD,Bresnahan JC.A sensitive and reliable locomotor rating scale for open field testing in rats[J].J Neurotrauma,1995,12(1):1
[9]胡爱民,张治国,侯志贞,等.褪黑素对大鼠实验性脊髓损伤的神经保护作用[J].解放军医学杂志,2011,36(2):164
[10]Wakabayashi K,Nagai A,Sheikh AM,et al.Transplantation of human mesenchymal stem cells promotes functional improvement and increased expression of neurotrophic factors in a rat focal cerebral ischemia model[J].J Neurosci Res,2010,88(5):1017
[11]Li J,Lepski G.Cell transplantation for spinal cord injury:a systematic review[J].Biomed Res Int,2013,2013:786475
[12]Dai G,Liu X,Zhang Z,et al.Transplantation of autologous bone marrow mesenchymal stem cells in the treatment of complete and chronic cervical spinal cord injury[J].Brain Res,2013,1533:73
[13]张群,王竞秋,宋焱峰,等.大鼠急性脊髓损伤后MCP-1及BDNF表达变化的实验研究[J].甘肃医药,2013,32(3):184
[14]徐玉生,张松,金伟林,等.褪黑素对大鼠脊髓损伤后肿瘤坏死因子-α表达的影响[J].郑州大学学报:医学版,2014,49(1):41