【摘要】 目的:观察核苷酸类神经营养因子三磷酸腺苷对大鼠脊髓神经元损伤后细胞活力的影响。方法:取新生Wistar大鼠20只,切取脊髓,剪碎组织成糜状进行神经细胞培养。将培养的神经细胞分为3组,①无损伤对照组,继续完全培养液培养。②损伤组,培养第7天神经元用微量移液器塑料滴头行机械性划痕损伤后完全培养液培养。③三磷酸腺苷组,培养第7天神经元划痕损伤后加入终浓度为1.0mmol/L的三磷酸腺苷培养液培养。倒置相差显微镜观察细胞生长及不同时期细胞形态变化,各组于处理后10min,1,12,24和48h,细胞损伤程度,以乳酸脱氢酶法检测(乳酸脱氢酶渗漏量比值越大,说明细胞损伤越重)。检测各组细胞活性采用四甲基偶氮唑盐比色法(四甲基偶氮唑盐代谢率吸光度越高,说明细胞活性越高)。结果:①各组细胞损伤程度评估:以培养液中乳酸脱氢酶含量表示。伤后1,12,24和48h,损伤组和三磷酸腺苷组较对照组明显增加(P<0.05);损伤24和48h,三磷酸腺苷组低于损伤组(17.94±0.82,23.05±1.04;18.52±1.12,24.88±1.16;P<0.05)。②体外培养的各组大鼠损伤神经元的变化:以四甲基偶氮唑盐代谢率表示。伤后1,12,24和48h,损伤组和三磷酸腺苷组明显低于对照组(P<0.05),损伤24和48h,三磷酸腺苷组高于损伤组(0.24±0.03,0.18±0.04;0.27±0.03,0.15±0.05;P<0.05)。结论:细胞外三磷酸腺苷可以减轻机械性损伤后的脊髓神经元的损伤程度,增强细胞活力,对受损伤的神经元有一定的保护作用。更多还原

【Abstract】 AIM:To study the effects of adenosine triphosphate on the activity of injured neurons in spinal cord of neonatal rats. METHODS:A total of 20 neonatal rats were selected to obtain spinal cord, nerve cells in broken tissues were cultured.The cultured nerve cells were divided into three groups:①nerve cells in control group were cultured in culture medium continually;②after spinal neurons were cultured for 7 days, mechanical injury was caused by plastic tip in the injury group and then the cells were cultured in complete medium continually;③adenosine triphosphate was added into the cultured medium of adenosine triphosphate group with the final concentration of 1.0 mmol/L following mechanical injury.The convert phase microscope was used to observe the growth and morphologic change of neurons.Cell viability and cell injury were detected by mononuclear cell direct cytotoxicity assay method (the higher absorbency of metabolic rate indicated the higher activity of cells) and lactate dehydrogenase assay(the higher ratio of lactate dehydrogenase leakage indicated the more serious injury) at 10 minutes,1,12,24 and 48 hours after injury.RESULTS:①The injured degree of cells: the content of lactate dehydrogenase increased in injury and adenosine triphosphate groups at 1,12,24 and 48 hours after injury as compared with that in control group(P< 0.05), which was decreased in adenosine triphosphate group at 24 and 48 hours as compared with that in injury group(17.94±0.82,23.05±1.04;18.52±1.12,24.88±1.16;P< 0.05).②The changes of injured neurons cultured in vitro: the metabolic rate decreased significantly in injury and adenosine triphosphate groups at 1,12,24 and 48 hours after injury as compared with that in control group(P< 0.05),which was increased in adenosine triphosphate group at 24 and 48 hours as compared with that in injury group(0.24±0.03,0.18±0.04;0.27±0.03,0.15±0.05;P< 0.05).CONCLUSION:Adenosine triphosphate in vitro can relieve the injured degree of spinal cord neruons,increase the activity of cells and have the protective effects on the injured spinal neurons of neonatal rats.更多还原