火针调控脊髓损伤后神经干细胞Ngn1、CyclinD1基因表达的离体研究
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摘要
目的:观察火针对脊髓损伤后神经干细胞Ngn1和Cyclin D1基因的调控作用。方法:采用随机数字表法将60只大鼠随机分为空白组(5只)、假手术组(5只),剩余大鼠按照1天、3天、7天、10天、14天时间点随机分为模型组、火针组,每组5只;采用改良的Allen’s法对模型组和火针组造模,相应时间点取大鼠腹主动脉血清置于神经干细胞培养皿中培养72 h,通过RT-PCR技术检测目的基因表达。结果:与假手术组比较,1天时模型组和火针组Ngn1基因表达量明显降低,差异具有统计学意义(P<0.05);在7天、10天时火针组Ngn1基因表达量均明显高于模型组,差异具有统计学意义(P<0.05)。与假手术组比较,1天时模型组和火针组Cyclin D1基因表达量明显降低,差异具有统计学意义(P<0.05);在3天、7天、10天时火针组Cyclin D1基因表达量均明显高于模型组,差异具有统计学意义(P<0.05)。结论:火针可调控脊髓损伤神经干细胞Ngn1和Cyclin D1基因表达。
Objective:To explore the regulatory effect of fire needling on genes Ngn1 and Cyclin D1 of neural stem cells(NSCs) in rats with spinal cord injury(SCI).Methods:60 SD rats were randomized into the blank group(n = 5),sham operation group(n = 5),and the rest were randomized into the model group(n = 25) and the fire needling group(n = 25),according to time points of 1 d,3 d,7 d,10 d,and 14 d.Modified Allen's method was applied to build SCI model;RT-PCR was applied to detect the genes after 72 hours of NSCs culture.Results:The expression of Ngn1 gene in the model group and the fire needling group significantly decreased compared to the sham operation group on 1st day(P < 0.05);and it was significantly higher in the fire needling group than that in the model group on 7th and 10 th day(P < 0.05).The expression of Cyclin D1 gene in the model group and the fire needling group significantly decreased compared to sham operation group on 1st day(P < 0.05);and it was significantly higher in the fire needling group compared to that in the model group on 3rd,7th and 10 th day(P < 0.05).Conclusion:Fire needling could modulate the gene expressions of Ngn1 and Cyclin D1 in neural stem cells.
Objective:To explore the regulatory effect of fire needling on genes Ngn1 and Cyclin D1 of neural stem cells(NSCs) in rats with spinal cord injury(SCI).Methods:60 SD rats were randomized into the blank group(n = 5),sham operation group(n = 5),and the rest were randomized into the model group(n = 25) and the fire needling group(n = 25),according to time points of 1 d,3 d,7 d,10 d,and 14 d.Modified Allen's method was applied to build SCI model;RT-PCR was applied to detect the genes after 72 hours of NSCs culture.Results:The expression of Ngn1 gene in the model group and the fire needling group significantly decreased compared to the sham operation group on 1st day(P < 0.05);and it was significantly higher in the fire needling group than that in the model group on 7th and 10 th day(P < 0.05).The expression of Cyclin D1 gene in the model group and the fire needling group significantly decreased compared to sham operation group on 1st day(P < 0.05);and it was significantly higher in the fire needling group compared to that in the model group on 3rd,7th and 10 th day(P < 0.05).Conclusion:Fire needling could modulate the gene expressions of Ngn1 and Cyclin D1 in neural stem cells.
引文
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[17]胡爱娥.膀胱三角区针刺治疗脊髓损伤患者排尿功能障碍的效果[J].针灸临床杂志,2015,31(7):34-36
[18]张晓梅,高维滨,滕秀英,等.夹脊电针及预处理对大鼠脊髓损伤后内源性神经干细胞的影响[J].针灸临床杂志,2011,27(11):53-56
[19]胡科,任应国,徐明超,等.针刺联合康复训练对MCAO大鼠内源性神经干细胞增殖的影响[J].南京中医药大学学报,2016,32(1):50-53
[2]Lepore AC,Walczak P,Rao MS,et al.MR imaging of lineage-restricted neural precursors following transplantation into the adult spinal cord[J].Experimental neurology,2006,201(1):49-59
[3]Reier PJ.Cellular transplantation strategies for spinal cord injury and translational neurobiology[J].NeuroRx:the journal of the American Society for Experimental Neuro Therapeutics,2004,1(4):424-51
[4]孙立明,李岩,周震,等.火针对脊髓损伤模型大鼠凋亡相关蛋白表达的影响[J].针灸临床杂志,2010,26(12):49-53
[5]Cristante AF,Barros Filho TE,Marcon RM,et al.Therapeutic approaches for spinal cord injury[J].Clinics,2012,67(10):1219-24
[6]Hyun JK,Kim HW.Clinical and experimental advances in regeneration of spinal cord injury[J].Journal of tissue engineering,2010,2010:650857
[7]Bamford JA,Mushahwar VK.Intraspinal microstimulation for the recovery of function following spinal cord injury[J].Progress in brain research,2011,194:227-39
[8]张月兰,罗莎,田松焕.中药熏蒸对脊髓损伤患者下肢肌张力及日常生活活动能力的影响[J].中华物理医学与康复杂志,2016,38(7):541-543
[9]吕春风,李京,曹锐,等.高压氧联合中药“蝎七方”改善急性脊髓损伤患者神经功能的临床观察[J].中华航海医学与高气压医学杂志,2015,22(6):437-440
[10]Duan H,Song W,Zhao W,et al.Endogenous neurogenesis in adult mammals after spinal cord injury[J].Science China Life sciences,2016,59(12):1313-8
[11]苑文超,袁翰,郭杨,等.诱导多能干细胞修复脊髓损伤的机制研究与应用进展[J].广东医学,2016,37(21):3286-3289
[12]马娇,张雪梅.骨髓间充质干细胞治疗脊髓损伤的分子机制[J].脑与神经疾病杂志,2016,24(2):117-119
[13]Zhao Y,Zuo Y,Jiang J,et al.Neural stem cell transplantation combined with erythropoietin for the treatment of spinal cord injury in rats[J].Experimental and therapeutic medicine,2016,12(4):2688-94
[14]Yan GZ,Ziff EB.NGF regulates the PC12 cell cycle machinery through specific inhibition of the Cdk kinases and induction of cyclin D1[J].The Journal of neuroscience:the official journal of the Society for Neuroscience,1995,15(9):6200-12
[15]Megason SG,Mc Mahon AP.A mitogen gradient of dorsal midline Wnts organizes growth in the CNS[J].Development,2002,129(9):2087-98
[16]王德强.针刺治疗脊髓损伤所致排尿功能障碍的疗效观察[J].中国医学创新,2015,12(34):91-93
[17]胡爱娥.膀胱三角区针刺治疗脊髓损伤患者排尿功能障碍的效果[J].针灸临床杂志,2015,31(7):34-36
[18]张晓梅,高维滨,滕秀英,等.夹脊电针及预处理对大鼠脊髓损伤后内源性神经干细胞的影响[J].针灸临床杂志,2011,27(11):53-56
[19]胡科,任应国,徐明超,等.针刺联合康复训练对MCAO大鼠内源性神经干细胞增殖的影响[J].南京中医药大学学报,2016,32(1):50-53