电针干预脊髓损伤大鼠受损节段缺氧诱导因子1α、血管内皮生长因子的表达

背景:作者发现,中医“痿病”从气血论治理论与脊髓损伤神经修复的关键环节在于改善组织血氧微环境有着惊人的相似,提出假设:电针刺激可通过干预缺氧诱导因子1α、血管内皮生长因子信号转导,改善脊髓血氧微环境从而促进神经再生。目的:探讨电针干预对脊髓损伤大鼠受损节段缺氧诱导因子1α、血管内皮生长因子表达的影响。方法:120只SD雌性大鼠以微型血管夹夹闭脊髓,保持夹闭状态20 s制作脊髓夹伤模型,随机分为3组:即阿是穴电针组、足阳明胃经电针组和空白对照组,每组40只。电针组造模后第3天开始接受每日1次的电针治疗,阿是穴电针组选择2个阿是穴、足阳明胃经电针组选择双侧伏兔、足三里穴,每周5次。于干预后1,2,3,4,5周对大鼠进行BBB评分之后分别取出损伤的脊髓组织标本,行病理组织观察,并采用免疫组织化学染色、实时荧光定量PCR技术及Western blot检测损伤脊髓缺氧诱导因子1α、血管内皮生长因子的基因和蛋白表达。实验方案经广西中医药大学第一附属医院动物实验伦理委员会批准(批准号201712001)。结果与结论:①阿是穴电针组及足阳明胃经电针组大鼠的下肢功能评分、缺氧诱导因子1α及血管内皮生长因子基因及蛋白表达明显高于未经电针干预的对照组;②随着干预时间的推移阿是穴电针组及足阳明胃经电针组的神经元数量均明显高于空白对照组;③结果说明,电针干预可以有效改善脊髓损伤大鼠的下肢功能评分、增加神经元的数量、上调缺氧诱导因子1α及血管内皮生长因子的mRNA及蛋白表达,从而有效促进脊髓神经功能的修复。

Effect of electroacupuncture intervention on expression of hypoxia-inducible factor-1 alpha and vascular endothelial growth factor in injured segments of rats with spinal cord injury

BACKGROUND:The authors found a striking similarity between the qi and blood theory and nerve repair of spinal cord injury in terms of improving blood-oxygen microenvironment in tissues.The hypothesis is that electroacupuncture can improve the blood-oxygen microenvironment of the spinal cord and promote nerve regeneration by regulating hypoxia-inducible factor 1a and vascular endothelial growth factor signal transduction.OBJECTIVE:To investigate the effect of electroacupuncture intervention on the expression of hypoxia-inducible factor 1a and vascular endothelial growth factor in injured segments of spinal cord injury rats.METHODS:Totally 120 Sprague-Dawley female rats were enrolled to make spinal cord injury models by clamping the spinal cord(20 seconds)using a microvascular clamp.Rat models were then randomly divided into three groups:Ashi point group,Stomach Meridian of Foot-Yangming group and blank control group,40 rats in each group.Electroacupuncture at two Aishi points or at both sides of Futu and Zusanli points was started on the 3rd day after modeling.Each rat was scored on the Basso,Beattie and Bresnahan locomotor rating scale at 1,2,3,4,and 5 weeks after intervention.Injured spinal cord specimens were then taken and observed histomorphologically.Hypoxia-inducible factor-1a and vascular endothelial growth factor protein and mRNA expressions were detected using immunohistochemistry staining,real-time fluorescence quantitative PCR and western blot assay.The study protocol was approved by the Animal Ethic Committee of the First Affiliated Hospital of Guangxi University of Chinese Medicine in China(approval No.201712001).RESULTS AND CONCLUSION:The lower limb function score,hypoxia-inducible factor-1a and vascular endothelial growth factor gene and protein expression in the two electroacupuncture intervention groups were significantly higher than those of the blank control group.The number of neurons in Ashi point and Stomach Meridian of Foot-Yangming groups was significantly higher than that of the blank control group with the lapse of intervention time.Electroacupuncture intervention can effectively improve the lower limb function score of spinal cord injury rats,increase the number of neurons,and up-regulate the mRNA and protein expression of hypoxia-inducible factor-1a and vascular endothelial growth factor,thus effectively promoting the neurological recovery of the spinal cord.