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[艾灸对血管性痴呆大鼠额叶皮质和海马中RAGE和LRP-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].

作者信息

Wang Jing Ji, Wu Sheng Bing, Cheng Hong Liang, Yang Chao, Chen Yun, Yang Jun

机构信息

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.

出版信息

Zhen Ci Yan Jiu. 2020 Jan 25;45(1):33-9. doi: 10.13702/j.1000-0607.1902956.

Abstract

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 (=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 day of modeling (<0.01). The expression levels of RAGE protein and mRNA and APP mRNA were significantly increased (<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 (<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 (<0.01). Results of electron microscopy showed dilation of the mitochondria with disappearance of the cristae, partial vacuolar degeneration or dissolved external membrane, and cytoplasmic edema with basic disappearance of the rough endoplasmic reticulum and glycogen particles in neurons of the frontal cortex and hippocampal CA1 region after modeling, which was relatively milder in the moxibustion group.

CONCLUSION

Moxibustion of acupoints of the Governor Vessel may reduce the content of APP in the frontal cortex and hippocampus by regulating the expression of RAGE and LRP-1 proteins and mRNA in VD rats.

摘要

目的

探讨督脉穴位艾灸对血管性痴呆(VD)大鼠学习记忆能力、晚期糖基化终末产物受体(RAGE)、低密度脂蛋白受体相关蛋白1(LRP-1)、β淀粉样前体蛋白(APP)表达及额叶皮质和海马神经元超微结构的影响,以探讨其缓解VD的潜在机制。

方法

将24只雄性SD大鼠随机分为正常组、模型组和艾灸组(每组8只)。采用双侧颈总动脉阻断20分钟再灌注10分钟,重复3次的方法建立VD模型。艾灸“百会”(GV20)、“大椎”(GV14)和“风府”(GV16)30分钟,每日1次,共4周。采用Morris水迷宫试验评估学习记忆能力。实验结束时,取大脑皮质额叶和海马组织,分别用蛋白质免疫印迹法检测RAGE和LRP-1蛋白表达,用实时荧光定量PCR检测RAGE、LRP-1和APP mRNA表达。用透射电子显微镜观察额叶皮质和海马CA1区神经元的超微结构。

结果

造模后,Morris水迷宫试验显示,造模第4天模型组和艾灸组平均逃避潜伏期均显著延长(<0.01)。模型组与正常组相比,皮质和海马中RAGE蛋白和mRNA以及APP mRNA表达水平显著升高(<0.01);而LRP-1蛋白和mRNA表达显著降低(<0.01)。艾灸后,艾灸组与模型组相比,造模诱导的RAGE蛋白和mRNA以及APP mRNA表达升高以及LRP-1蛋白和mRNA表达下调得到逆转(<0.01)。电镜结果显示,造模后额叶皮质和海马CA1区神经元线粒体肿胀、嵴消失;部分空泡变性或外膜溶解;细胞质水肿,粗面内质网和糖原颗粒基本消失,艾灸组上述改变相对较轻。

结论

督脉穴位艾灸可能通过调节VD大鼠RAGE和LRP-1蛋白及mRNA表达,降低额叶皮质和海马中APP含量。

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