Effect of the warming kidney for dredging governor channel acupuncture method on related neurotransmitters and the cAMP-RAF1 signal pathway on depression with kidney yang deficiency syndrome model rats

CAO Lingxiu; REN Lu

doi:10.3969/j.issn.1006-2157.2023.01.023

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Effect of the warming kidney for dredging governor channel acupuncture method on related neurotransmitters and the cAMP-RAF1 signal pathway on depression with kidney yang deficiency syndrome model rats

Acupuncture Moxibustion and Tuina | 更新时间：2023-02-03

- Effect of the warming kidney for dredging governor channel acupuncture method on related neurotransmitters and the cAMP-RAF1 signal pathway on depression with kidney yang deficiency syndrome model rats
- Journal of Beijing University of Traditional Chinese Medicine Vol. 46, Issue 1, Pages: 141-148(2023)
- 作者机构：
  
  1.辽宁中医药大学　沈阳　110847
  2.郑州大学第五附属医院康复医学科
- 作者简介：
  
  Prof. REN Lu, Ph.D., Doctoral Supervisor. Liaoning University of Traditional Chinese Medicine, No.79, Chongshan Donglu Road, Huanggu District, Shenyang 110847. E-mail: renlu2008.student@sina.com
- 基金信息：
  
  National Key Research and Development Plan of China(2018YFC1704300);National Natural Science Foundation of China(82274665);Key R&D and Promotion Project of Henan Province(222102310491);Innovation Team of Colleges and Universities of Liaoning Province(辽教函〔2018〕479号);Special Professor Program of Liaoning Province(辽教发〔2015〕153号)
- DOI：10.3969/j.issn.1006-2157.2023.01.023
  CLC： R245
- Received：15 October 2021，
  
  Published Online：08 October 2022，
  
  Published：30 January 2023
- 稿件说明：
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CAO Lingxiu, REN Lu. Effect of the warming kidney for dredging governor channel acupuncture method on related neurotransmitters and the cAMP-RAF1 signal pathway on depression with kidney yang deficiency syndrome model rats[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(1): 141-148.
DOI：

CAO Lingxiu, REN Lu. Effect of the warming kidney for dredging governor channel acupuncture method on related neurotransmitters and the cAMP-RAF1 signal pathway on depression with kidney yang deficiency syndrome model rats[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(1): 141-148. DOI： 10.3969/j.issn.1006-2157.2023.01.023.

摘要

目的

通过建立抑郁症肾阳虚证模型大鼠，探究温肾通督针法的温肾调神功效，揭示温肾通督针法温阳解郁的作用机制。

方法

将大鼠任意分为正常组、模型组、温针组、抑制剂组、温针＋抑制剂组，每组各6只。除正常组外，其余各组采用氢化可的松激素注射(肌肉注射，25 mg/kg)结合慢性不可预见性温和刺激进行造模。温针组和温针＋抑制剂组采用针刺和艾灸2种方式进行干预；温针＋抑制剂组和抑制剂组在取材前24 h灌胃给药ESI－09[10 mg/kg，环磷酸腺苷(cAMP)－RAF1信号通路抑制剂]。造模及干预前后，观察大鼠一般情况，采用强迫游泳实验、水迷宫实验观察大鼠行为学变化。酶联免疫吸附测定法检测脑源性神经营养因子(BDNF)、5－羟色胺(5－HT)、多巴胺(DA)及去甲肾上腺素(NE)含量。蛋白质印迹法检测大鼠海马cAMP-RAF1信号通路中cAMP、RAS相关蛋白1(RAP1)和RAF1的蛋白表达。

结果

干预后，与正常组比较，模型组、温针组、抑制剂组和温针＋抑制剂组的体质量均降低(

0.01)，模型组、抑制剂组的肛温均降低(

0.01)。与模型组比较，抑制剂组的体质量、肛温降低(

0.05，

0.01)，温针组、温针＋抑制剂组的体质量、肛温升高(

0.05，

0.01)。干预后，与正常组比较，模型组、抑制剂组和温针＋抑制剂组的强迫游泳不动时间均延长(

0.01)，温针组强迫游泳不动时间差异无统计学意义。与模型组比较，抑制剂组强迫游泳不动时间延长(

0.01)，温针组、温针＋抑制剂组强迫游泳不动时间均缩短(

0.05，

0.01)。干预后，与正常组比较，模型组、温针组、抑制剂组和温针＋抑制剂组平台潜伏期、平台区域穿梭次数、平台象限时间占比均降低(

0.01)。与模型组比较，抑制剂组平台潜伏期、平台区域穿梭次数、平台象限时间占比均降低(

0.01)，温针组、温针＋抑制剂组平台潜伏期、平台区域穿梭次数、平台象限时间占比均升高(

0.01)。模型组大鼠BDNF、5－HT、DA、NE含量及海马cAMP、RAP1、RAF1蛋白表达均低于正常组(

0.01)，温针组大鼠BDNF、5－HT、DA、NE含量及海马cAMP、RAP1、RAF1蛋白表达均高于模型组(

0.01)。

结论

温肾通督针法可能通过调控cAMP-RAF1信号通路，使海马功能得以恢复，从而改善大鼠抑郁样行为。

Abstract

Objective

By estabtishing depression with kidney yang deficiency syndrome model rats

to explore the effect of warming kidney for dredging governor channel acupuncture method on warming kidney and regulating spirit

and reveal the mechanism underlying the warming kidney for dredging governor channel acupuncture method.

Methods

The rats were randomly divided into the normal group

model group

warm needle group

inhibitor group

and warm needle combined with inhibitor group

with six rats per group. Except for the normal group

the other groups were given a hydrocortisone hormone injection (intramuscular injection

25 mg/kg) combined with chronic unpredictable mild stimulation for modeling. Acupuncture and moxibustion were used in the warm needle group and warm needle combined with inhibitor group. The rats in the warm needle combined with inhibitor group and inhibitor group were orally treated with ESI-09 (cAMP-RAF1 signal pathway inhibitor) at a dose of 10 mg/kg 24 h before being sacrificed. Before and after modeling and intervention

the general conditions of rats were observed

and forced swimming and water maze tests were used to observe the behavioral changes in rats. The levels of brain-derived neurotrophic factor (BDNF)

5-hydroxytryptophan (5-HT)

dopamine (DA)

and norepinephrine (NE) were determined by ELISA. Western blotting was used to detect the expression of cAMP

RAS-related protein 1 (RAP1)

and RAF1 proteins in the cAMP-RAF1 signal pathway.

Results

After intervention

compared with the normal group

the body weights of the model group

warm needle group

inhibitor group

and warm needle combined with inhibitor group were decreased (

0.01)

as were the anal temperatures of the model group and inhibitor group (

0.01). Compared with the model group

the body weight and anal temperature of the inhibitor group were decreased (

0.05

0.01)

while those of the warm needle group and warm needle combined with inhibitor group were increased (

0.05

0.01). After intervention

compared with the normal group

the forced swimming immobility time of the model group

the inhibitor group and the warm needle combined with inhibitor group was prolonged (

0.01)

and there was no significant difference in the forced swimming immobility time of the warm needle group. Compared with the model group

the forced swimming immobility time of the inhibitor group was prolonged (

0.01)

and the forced swimming immobility time of the warm needle group and warm needle combined with inhibitor group was shortened (

0.05

0.01). After intervention

compared with the normal group

the platform latency

shuttle times of the platform area

and the platform quadrant time ratio of the model group

warm needle group

inhibitor group and warm needle combined with inhibitor group were all decreased (

0.01). Compared with the model group

the platform latency

shuttle times of the platform area

and the platform quadrant time ratio of the inhibitor group were decreased (

0.01)

but were increased in the warm needle group and warm needle combined with inhibitor group (

0.01). The contents of BDNF

5-HT

and NE

and the protein expressions of cAMP

RAP1

and RAF1 in the hippocampus of the model group were lower than those of the normal group (

0.01)

but those were higher in the warm needle group compared with the model group (

0.01).

Conclusion

Warming kidney for dredging governor channel acupuncture may restore hippocampal function by regulating the cAMP-RAF1 signal pathway

thus improving depression-like behavior in rats.

关键词

Keywords

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