电针脊髓损伤模型大鼠“足三里”与“伏兔”穴神经细胞凋亡因子Caspase-3的表达  

Effect of Electroacupuncture at Zusanli and Futu acupoints on expression of apoptosis factor Caspase-3 in spinal cord injury rats

作　　者：魏卫兵 周宾宾[2] 张鸿升 杨宇[2] 崔俊武[2] 李振兴[2] Wei Weibing;Zhou Binbin;Zhang Hongsheng;Yang Yu;Cui Junwu;Li Zhenxing(Liuzhou Hospital of Traditional Chinese Medicine,Liuzhou 545000,Guangxi Zhuang Autonomous Region,China;First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530000,Guangxi Zhuang Autonomous Region,China;Ruikang Hospital,Guangxi University of Chinese Medicine,Nanning 530000,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]柳州市中医医院,广西壮族自治区柳州市545000 [2]广西中医药大学第一附属医院,广西壮族自治区南宁市530000 [3]广西中医药大学附属瑞康医院,广西壮族自治区南宁市530000

出　　处：《中国组织工程研究》2020年第32期5151-5157,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81660814),项目负责人:周宾宾;广西自然科学基金(2016JJA140159,2017JJA140554z),项目负责人:周宾宾。

摘　　要：背景:中医自《内经》开始就提出了"治痿独取阳明"之理论,但是有关电针干预足阳明胃经治疗脊髓损伤的报道相对较少。目的:探讨电针刺激"足三里""伏兔"穴对脊髓损伤大鼠受损节段Caspase-3表达的影响。方法:将64只SPF级SD雌性大鼠随机分为4组,即对照组、预标记组、损伤后标记组和电针干预组,每组16只。对照组及预标记在操作前给予Brdu(50mg/kg)连续10d腹腔注射标记细胞,对照组于第11天给予咬除椎板,并不损伤脊髓,预标记组第11天给予制作脊髓损伤模型;损伤后标记组及电针组在制作脊髓损伤后连续10 d腹腔注射Brdu标记细胞,其中损伤后标记组不给予干预措施,电针组造模后第3天开始针刺足三里、伏兔穴。各组分别于损伤后3,10,17,24 d取脊髓组织,运用苏木精-伊红染色观察神经元数量的变化;免疫组织化学染色观察Brdu阳性细胞表达;q RT-PCR技术检测Caspase-3mRNA的变化;Western blot检测Caspase-3蛋白表达的变化。实验方案经广西医科大学动物实验伦理委员会批准(批准号201712001)。结果与结论:①脊髓损伤以后Caspase-3的mRNA及蛋白表达均高于脊髓没有受到损伤的对照组,脊髓损伤后神经元的数量明显较对照组少;随着时间的推移,预标记组、损伤后标记组及电针干预组Caspase-3mRNA及蛋白表达先升高后降低,而电针组的Caspase-3表达明显减少,与预标记组及损伤后标记组比较差异有显著性意义(P<0.05);②苏木精-伊红染色显示随着时间的推移电针组的神经元数量明显比未经干预的预标记组、损伤后标记组高,在第24天时逐渐接近对照组的神经元数量;③免疫组织化学结果显示电针组阳性蛋白表达呈先升高后降低的趋势,17 d升至最高,24 d降至最低,整体阳性蛋白表达高于其他组(P<0.05);④结果说明,电针刺激"足三里""伏兔"穴可以减少Caspase-3 mRNA及蛋白表达,减少神经细胞的凋亡,同时可以促BACKGROUND:Traditional Chinese medicine has proposed the theory of"treatment for flaccidity aims at Yangming meridian"in the Internal Canon of Medicine.However,there have been relatively few reports on electroacupuncture at the stomach channel of Foot Yangming in the treatment of spinal cord injury.OBJECTIVE:To investigate the effect of electroacupuncture stimulation of Zusanli and Futu on Caspase-3 expression in injured segments of spinal cord injury rats.METHODS:Sixty-four Sprague-Dawley female rats,SPF grade,were randomly divided into four groups,i.e.,control group,pre-labeling group,post-injury labeling group and electroacupuncture group,with 16 rats in each group.The control group and the pre-labeling group were intraperitoneally given Brdu(50 mg/kg)for 10 days before the operation.The control group was given a bite to remove the lamina on the 11 th day,which did not damage the spinal cord.A model of spinal cord injury was made in the pre-labeling group on the 11 th day.The post-injury labeling group and the electroacupuncture group were injected intraperitoneally with Brdu for 10 days after the spinal cord injury was made,and the post-injury labeling group was not treated.The electroacupuncture group started electroacupuncture at Zusanli and Futu acupoints on the 3 rd day after the model was established.Spinal cord specimens were taken at 3,10,17,and 24 days after injury in each group,and the changes in the number of neurons were observed by hematoxylin-eosin staining.The expression of Brdu positive cells was observed by immunohistochemistry.The changes in Caspase-3 mRNA and protein expression were detected by qRT-PCR and western blot,respectively.The study protocol was approved by the Animal Ethic Committee of Guangxi Medical University,with an approval No.201712001.RESULTS AND CONCLUSION:The expression of Caspase-3 mRNA and protein after spinal cord injury was higher than that of the control group without spinal cord injury,and the number of neurons after spinal cord injury was significantly lower than th

关 键 词：脊髓损伤 电针刺激 足三里 伏兔 CASPASE-3 大鼠 实验 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]