神经干细胞移植数量对大鼠神经病理性痛的影响

目的 探讨神经干细胞(NSGs)移植数量对大鼠神经病理性痛的影响.方法 取出生1～3 d的SD大鼠,制备NSCs悬液.清洁级雄性SD大鼠84只,体重150～180 g,采用右侧坐骨神经半切断法制备大鼠神经病理性痛模型.采用随机数字表法,将大鼠随机分为7组(n=12),假手术组(S组):仅暴露坐骨神经,不切断,鞘内注射细胞培养液30 μl;神经病理性痛组(NP组):于模型制备后3d鞘内注射细胞培养液30 μl;NP+NSCs 103组(N1组)、NP+NSCs 104组(N2组)、NP+NSCs 105组(N3组)、NP+NSCs 106组(N4组)和NP+NSCs 107组(N5组):于模型制备后3 d,鞘内注射相应浓度的NSCs悬液.模型制备前1 d和制备后1、3.7、14和21 d时测右足机械缩足阈值(MWT)和热缩足潜伏期(TWL);于模型制备后7和21 d,痛阈测定结束后,取右侧腰膨大脊髓背角及L5背根神经节,测定脑源性神经营养因子(BDNF)mRNA及其蛋白的表达水平.结果 与S组比较,NP组和N1～5组MWT降低,TWL缩短,脊髓背角和DRG BDNF mRNA表达上调(P<0.05),BDNF阳性细胞数增多,染色加深;与NP组比较,N2～5组MWT升高,TWL延长,脊髓背角和DRG BDNF mRNA表达上调(P<0.05),BDNF 阳性细胞数增多,染色加深;模型制备后7 d N1～5组MWT依次升高,TWL依次延长,脊髓背角和DRG BDNF mRNA表达依次上调(P<0.05),BDNF阳性细胞数依次增多,染色逐渐加深;模型制备后14、21d,N1～3组MWT依次升高,TWL依次延长(P<0.05),模型制备后21 d,N1～3组脊髓背角和DRG BDNF mRNA表达依次上调,BDNF阳性细胞数依次增多,染色逐渐加深;N3～5组上述指标比较差异无统计学意义(P>0.05).结论 NSCs移植减轻大鼠神经病理性痛的适宜移植数量为105个.

Effect of intrathecal transplantation of different quantities of neural stem cells on neuropathic pain in rats

Objective To investigate the effect of intrathecal (IT) transplantation of different quantities of neural stem cells (NSCs) on neuropathic pain (NP) in rats.Methods Eighty-four adult pathogen-free male SD rats weighing 150-180 g were randomly divided into 7 groups ( n = 12 each) : group sham operation (S group) , NP group, NP+ NSCs 103 , 104 , 105 , 106 , 107 groups (N1-5 groups) . NP was induced by partial transection of right sciatic nerve. NSCs were transplanted into subarachnoid space in N1-5 groups. Paw withdrawal threshold to mechanical stimulation (MWT) and paw withdrawal latency to nociceptive thermal stimuli (TWL) were measured at 1 day before (baseline) and 1, 3, 7, 14 and 21 days after operation. Brain derived neurotrophic factor ( BDNF) expression in spinal dorsal horn and dorsal root ganglia (DRG) was detected by immuno-histochemistry and RT-PCR on the 7th and 21st day after operation. Results Partial transection of the sciatic nerve gradually reduced MWT and TWL after operation starting from day 1 until day 7 and 14 as compared with the baseline in group NP. IT NSC transplantation significantly increased MWT and TWL and expression of BDNF in spinal dorsal horn and DRG in a dese-dependent manner at day 7 after operation in N1-5 groups as compared with group NP. There were no significant differences in MWT and TWL and BDNF expression among N3, N4 and N5 groups at day 21 after operation.Conclusion The proper quantity of transplanted NSCs which are able to ameliorate NP induced by partial transection of sciatic nerve in rats is 105 .