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[艾灸对血管性痴呆大鼠学习记忆能力及海马脑源性神经营养因子/酪氨酸激酶受体B表达的影响]

[Effects of moxibustion on learning and memory ability and the hippocampal BDNF/TrkB expressions in the rats of vascular dementia].

作者信息

Zhang Yuan-Yuan, Zhang Xiao-Jun, Yang Jun, Fan Yin-Qiu, Cui Jing-Cheng, Wang Pin, Zhang Qing-Ping

机构信息

the First Affiliated Hospital of Anhui University of TCM, Hefei 230031, China.

Anhui Integrated Chinese and Western Medicine Hospital.

出版信息

Zhongguo Zhen Jiu. 2019 Jan 12;39(1):65-71. doi: 10.13703/j.0255-2930.2019.01.014.

Abstract

OBJECTIVE

To explore the effects of (resolving stasis, promoting collateral circulation) moxibustion on learning and memory ability and the expressions of brain derived neurotrophic factor (BDNF) and tyrosine kinase B (TrkB) in the rats of vascular dementia (VD) in the microenvironment of neurovascular niche.

METHODS

Using 2-vessel occlusion (2-VO), the VD rat models were duplicated. The neural stem cells (NSCs) labeled with lentiviral vector-mediated enhanced green fluorescent protein (EGFP) were co-cultured with endothelial progenitor cells (EPCs) to structure the NSCs + EPCs implant. The implant was transplanted into the lateral ventricle of VD rats and the VD rat models with neurovascular niche were established. In No.1 experiment, the successful-modeled rats were divided into 3 groups, i.e. a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 12 rats in each one. No any treatment was provided in the model group and the NSCs + EPCs blank group. The moxibustion therapy was adopted in the NSCs + EPCs moxibustion group, in which, the suspending moxibustion technique was applied to "Baihui" (GV 20), "Dazhui" (GV 14) and "Shenting" (GV 24), 20 min at each acupoint. The treatment was given once every day and a 14-day treatment was as one course. Totally, 3 courses of treatment were required. At the end of treatment, Morris water maze experiment was adopted to determine the learning and memory ability of the rats in each group. In the No.2 experiment, the model rats were divided into 3 groups, a NSCs + EPCs moxibustion group, a NSCs + EPCs blank group and a model group, 18 rats in each one. In each group, according to the durations of treatment, 3 subgroups were divided and 6 rats in each one. The intervention method was same as the No.1 experiment. Additionally, after corresponding treatment course, using perfusion, the brains were collected in each subgroup and the slices were frozen. BDNF/TrkB expressions were observed in the immunofluorescence test.

RESULTS

After treatment, in the NSCs + EPCs moxibustion group, the escape incubation was reduced, the time of the first running-cross platform was shortened and the frequency of running-cross platform increased as compared with the model group and the NSCs + EPCs blank group (<0.01, <0.05). The protein expressions were increased in tendency among the 3 courses of treatment in the NSCs + EPCs moxibustion group, indicating the significant differences (all <0.05), in which, the increase of the protein expressions in the NSCs + EPCs moxibustion group was better than the NSCs + EPCs blank group (<0.05, <0.01).

CONCLUSION

The moxibustion therapy is the effective approach to VD in clinical treatment. This therapy up-regulates the BDNF/TrkB protein expressions in the microenvironment of neurovascular niche, co-modulates NSCs-EPCs coupling mechanism, promotes nerve neogenesis and repairs the injured nerve.

摘要

目的

探讨化瘀通络艾灸对神经血管单元微环境中血管性痴呆(VD)大鼠学习记忆能力及脑源性神经营养因子(BDNF)和酪氨酸激酶B(TrkB)表达的影响。

方法

采用双血管阻断法(2-VO)复制VD大鼠模型。将慢病毒载体介导的增强型绿色荧光蛋白(EGFP)标记的神经干细胞(NSCs)与内皮祖细胞(EPCs)共培养构建NSCs+EPCs移植物。将移植物移植到VD大鼠侧脑室,建立具有神经血管单元的VD大鼠模型。实验一,将造模成功的大鼠分为3组,即NSCs+EPCs艾灸组、NSCs+EPCs空白组和模型组,每组12只。模型组和NSCs+EPCs空白组不做任何处理。NSCs+EPCs艾灸组采用艾灸疗法,采用悬灸技术灸“百会”(GV 20)、“大椎”(GV 14)和“神庭”(GV 24),每穴灸20分钟。每天治疗1次,14天为1个疗程,共治疗3个疗程。治疗结束后,采用Morris水迷宫实验检测各组大鼠的学习记忆能力。实验二,将模型大鼠分为3组,即NSCs+EPCs艾灸组、NSCs+EPCs空白组和模型组,每组18只。每组根据治疗时间分为3个亚组,每组6只。干预方法同实验一。此外,在相应疗程治疗后,每组亚组采用灌注法取脑并冻存切片,免疫荧光检测BDNF/TrkB表达。

结果

治疗后,NSCs+EPCs艾灸组与模型组和NSCs+EPCs空白组比较,逃避潜伏期缩短,首次穿越平台时间缩短,穿越平台频率增加(<0.01,<0.05)。NSCs+EPCs艾灸组在3个疗程治疗中蛋白表达呈上升趋势,差异有统计学意义(均<0.05),其中NSCs+EPCs艾灸组蛋白表达升高优于NSCs+EPCs空白组(<0.05,<0.01)。

结论

化瘀通络艾灸疗法是临床治疗VD的有效方法。该疗法上调神经血管单元微环境中BDNF/TrkB蛋白表达,协同调节NSCs-EPCs耦合机制,促进神经新生,修复受损神经。

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