艾灸督脉组穴对血管性痴呆大鼠皮层和海马中晚期糖基化终产物受体和低密度脂蛋白受体相关蛋白-1表达及神经元超微结构的影响  

Effect of moxibustion on expression of RAGE and LRP-1 and neuronal ultrastructure of frontal cortex and hippocampus in vascular dementia rats

作　　者：王婧吉[1,2] 吴生兵 程红亮 杨超[4] 陈赟 杨骏 WANG Jing-ji;WU Sheng-bing;CHENG Hong-liang;YANG Chao;CHEN Yun;YANG Jun(The Second Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230061,China;Institute of Acupuncture and Moxibustion,Anhui Academy of Chinese Medicine,Hefei 230061;Research and Experimental Center of Anhui University of Chinese Medicine,Hefei 230038;Graduate Faculty of Anhui University of Chinese Medicine,Hefei 230038;The First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230038)

机构地区：[1]安徽中医药大学第二附属医院,合肥230061 [2]安徽省中医药科学院针灸临床研究所,合肥230061 [3]安徽中医药大学科研实验中心,合肥230038 [4]安徽中医药大学研究生部,合肥230038 [5]安徽中医药大学第一附属医院,合肥230038

出　　处：《针刺研究》2020年第1期33-39,共7页Acupuncture Research

基　　金：安徽省自然科学基金项目(No.1608085QH220)

摘　　要：目的:观察艾灸督脉组穴对血管性痴呆(VD)大鼠大脑皮层和海马中晚期糖基化终产物受体(RAGE)、低密度脂蛋白受体相关蛋白-1(LRP-1)、β淀粉样蛋白前体(APP)表达和神经元超微结构的影响,探讨艾灸治疗VD的作用机制。方法:SD大鼠随机分为正常组、模型组、艾灸组,每组8只。采用结扎颈总动脉反复缺血再灌注法制备VD模型,艾灸组取"百会""大椎""风府"温和灸30min,每天1次,7d为1个疗程,共治疗4个疗程。采用Morris水迷宫法检测大鼠行为学变化,Western blot法检测各组大鼠大脑皮层和海马组织RAGE和LRP-1蛋白表达水平,实时荧光定量PCR法检测各组大鼠皮层和海马组织RAGE、LRP-1、APP mRNA表达水平,透射电镜观察大鼠大脑皮层额叶和海马CA1区超微结构。结果:Morris水迷宫结果显示,造模后第4天,模型组和艾灸组大鼠的平均逃避潜伏期较正常组显著延长(P<0.01)。治疗结束后,与正常组比较,模型组大鼠大脑皮层和海马组织中RAGE蛋白和mRNA表达显著升高(P<0.01),LRP-1蛋白和mRNA表达显著降低(P<0.01),APP mRNA表达明显升高(P<0.01)。与模型组比较,艾灸组大鼠大脑皮层和海马组织中RAGE蛋白和mRNA表达显著降低(P<0.01),LRP-1蛋白和mRNA表达显著升高(P<0.01),APP mRNA表达明显降低(P<0.01)。电镜结果显示,模型组皮层额叶和海马CA1区神经元线粒体明显扩张,外膜破坏、溶解,嵴排列紊乱或消失,胞质内水肿明显,糖原颗粒基本消失;艾灸组皮层额叶和海马CA1区神经元线粒体少数肿大、扩张,粗面内质网少数轻度扩张,糖原颗粒可见。结论:艾灸督脉组穴可能通过调节RAGE和LRP-1的表达水平降低脑内APP含量,改善VD大鼠大脑皮层额叶和海马CA1区神经元线粒体损伤。Objective To explore the effect of moxibustion of acupoints of Governor Vessel on learning-memory ability and expression of the receptor for advanced glycation end products(RAGE),low density lipoprotein(LDL)receptor related protein-1(LRP-1),β-amyloid precursor protein(APP)and neuronal ultrastructure of the frontal cortex and hippocampus in vascular dementia(VD)rats,so as to investigate its underlying mechanisms in relieving VD.Methods A total of 24 male SD rats were randomized into normal,model,and moxibustion groups(n=8 rats in each group).The VD model was established by occlusion of the bilateral common carotid arteries for 20 min and reperfusion for 10 min which was repeated for 3 times.Moxibustion was applied to"Baihui"(GV20),"Dazhui"(GV14)and"Fengfu"(GV16)for 30 min,once daily for 4 weeks.The learning-memory ability was evaluated by using Morris water maze tests.At the end of experiments,the frontal lobe of cerebral cortex and hippocampus tissues were collected for detecting the expression of RAGE and LRP-1 proteins with Western blot,and RAGE,LRP-1 and APP mRNAs with quantitative real-time PCR,respectively.The ultrastructure of neurons in the frontal cortex and hippocampal CA1 region was observed with transmission electron microscopy.Results Following modeling,Morris water maze tests showed that the average escape latency of the model group and moxibustion group was significantly prolonged on the 4 th day of modeling(P<0.01).The expression levels of RAGE protein and mRNA and APP mRNA were significantly increased(P<0.01),while those of LRP-1 protein and mRNA in the cortex and hippocampus remarkably decreased in the model group relevant to the normal group(P<0.01).Following moxibustion,modeling-induced increase of expression of RAGE protein and mRNA and APP mRNA,and down-regulation of expression of LRP-1 protein and mRNA were reversed in the moxibustion group relevant to the model group(P<0.01).Results of electron microscopy showed dilation of the mitochondria with disappearance of the cristae,partial vacuola

关 键 词：艾灸 血管性痴呆 晚期糖基化终产物受体 低密度脂蛋白受体相关蛋白-1 β淀粉样蛋白前体 超微结构 

分 类 号：R245.82[医药卫生—针灸推拿学]