电针“合谷”“太冲”对抑郁大鼠海马组织SLC6A4基因启动子区DNA甲基化的影响

张熙; 粟胜勇; 李欣; 王甜

doi:10.13703/j.0255-2930.20240902-k0005

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电针“合谷”“太冲”对抑郁大鼠海马组织SLC6A4基因启动子区DNA甲基化的影响

机制探讨 | 更新时间：2025-11-20

- 电针“合谷”“太冲”对抑郁大鼠海马组织SLC6A4基因启动子区DNA甲基化的影响
- Effect of electroacupuncture at "Hegu" (LI4) and "Taichong" (LR3) on DNA methylation of the SLC6A4 gene promoter in the hippocampus of depressed rats
- 中国针灸 2025年45卷第11期页码：1609-1616
- 作者机构：
  
  广西中医药大学研究生院，南宁　530001
  广西中医药大学第一附属医院针灸科，南宁　530023
  广西中医药防治医学分子生物重点实验室，南宁　530023
- 作者简介：
  
  ✉粟胜勇，主任医师。E-mail：1037097555@qq.com
- 基金信息：
  
  广西自然科学基金重点项目(2023GXNSFDA026052);广西中医药大学校级科研项目(2022QN014);广西研究生教育创新计划项目(2225240045)
- DOI：10.13703/j.0255-2930.20240902-k0005
  中图分类号：
- 收稿：2024-09-02，
  
  网络首发：2025-08-25，
  
  纸质出版：2025-11-12
- 稿件说明：
移动端阅览
张熙, 粟胜勇, 李欣, 等. 电针“合谷”“太冲”对抑郁大鼠海马组织SLC6A4基因启动子区DNA甲基化的影响[J]. 中国针灸, 2025,45(11):1609-1616.

ZHANG Xi, SU Shengyong, LI Xin, et al. Effect of electroacupuncture at "Hegu" (LI4) and "Taichong" (LR3) on DNA methylation of the SLC6A4 gene promoter in the hippocampus of depressed rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(11): 1609-1616.
张熙, 粟胜勇, 李欣, 等. 电针“合谷”“太冲”对抑郁大鼠海马组织SLC6A4基因启动子区DNA甲基化的影响[J]. 中国针灸, 2025,45(11):1609-1616. DOI： 10.13703/j.0255-2930.20240902-k0005.

ZHANG Xi, SU Shengyong, LI Xin, et al. Effect of electroacupuncture at "Hegu" (LI4) and "Taichong" (LR3) on DNA methylation of the SLC6A4 gene promoter in the hippocampus of depressed rats[J]. Chinese Acupuncture & Moxibustion, 2025, 45(11): 1609-1616. DOI： 10.13703/j.0255-2930.20240902-k0005.

摘要

目的：

观察电针“合谷”“太冲”对抑郁模型大鼠海马组织血清素再摄取转运体编码基因——溶质载体家族6成员4（SLC6A4）基因启动子区DNA甲基化的影响，探讨电针抗抑郁的效应机制。

方法：

将30只雄性SD大鼠随机分为空白组、模型组、西药组、5-氮杂胞苷（5-AZA）组和电针组，每组6只。模型组、西药组、5-AZA组和电针组采用慢性不可预知温和刺激（CUMS）结合孤养法构建抑郁大鼠模型。西药组予盐酸氟西汀胶囊灌胃；5-AZA组予腹腔注射5-AZA；电针组电针双侧“合谷”“太冲”，疏密波，频率2 Hz/100 Hz，电流1～1.2 mA，每次20 min。3组均每日干预1次，连续干预21 d。采用糖水偏好实验、旷场实验及新环境抑制进食实验评估行为学变化，采用ELISA法检测大鼠血清5-羟色胺（5-HT）、多巴胺（DA）、去甲肾上腺素（NE）含量；Western blot法、实时荧光定量PCR法检测大鼠海马组织SLC6A4、5-羟色胺1A受体（5-HT1AR）蛋白及mRNA表达；重亚硫酸氢盐测序（BSP）法检测大鼠海马组织SLC6A4基因启动子区DNA甲基化阳性率。

结果：

与空白组比较，模型组大鼠糖水偏好率降低、总行走距离减少、延迟进食时间延长（

0.05），血清5-HT、DA、NE含量降低（

0.05），海马组织SLC6A4、5-HT1AR蛋白及 mRNA 表达降低（

0.05），海马组织SLC6A4基因启动子区CpG位点甲基化阳性率升高（

0.05）。与模型组比较，西药组、5-AZA组、电针组糖水偏好率升高、总行走距离增加、延迟进食时间缩短（

0.05），血清5-HT、DA、NE含量升高（

0.05），海马组织SLC6A4、5-HT1AR蛋白及mRNA表达升高（

0.05），海马组织SLC6A4基因启动子区CpG位点甲基化阳性率降低（

0.05）。与西药组和5-AZA组比较，电针组大鼠糖水偏好率升高、总行走距离增加、延迟进食时间缩短（

0.05），血清DA、NE含量升高（

0.05）。

结论：

电针能够改善抑郁模型大鼠的抑郁行为，其机制可能是通过抑制SLC6A4基因高甲基化水平作用于5-HT能系统，上调SLC6A4、5-HT1AR蛋白及mRNA表达，使5-HT等单胺类神经递质含量升高。

Abstract

Objective

To observe the effect of electroacupuncture (EA) at "Hegu" (LI4) and "Taichong" (LR3) on DNA methylation of the solute carrier family 6 member 4 (SLC6A4) gene promoter region in the hippocampus of depressed rats

and to explore the potential antidepressant mechanism of EA.

Methods

Thirty male Sprague-Dawley rats were randomly divided into a blank group

a model group

a medication group

a 5-Azacytidine (5-AZA) group

and an EA group

6 rats in each group. Depression models were established in the model group

the medication group

the 5-AZA group

and the EA group using chronic unpredictable mild stress (CUMS) combined with solitary housing. The medication group was treated with intragastric administration of fluoxetine hydrochloride capsules; the 5-AZA group was treated with intraperitoneal injection of 5-AZA; the EA group was treated with EA at bilateral "Hegu" (LI4) and "Taichong" (LR3)

with disperse-dense wave

frequency of 2 Hz/100 Hz

and intensity of 1-1.2 mA

20 min each session. All the treatment was given in three groups once daily for 21 consecutive days. Behavioral changes were evaluated by sucrose preference test

open field test

and novelty-suppressed feeding test. Serum levels of serotonin (5-HT)

dopamine (DA)

and norepinephrine (NE) were measured by ELISA. The expression of SLC6A4 and 5-HT1AR protein and mRNA in hippocampus was detected by Western blot and real-time quantitative PCR

respectively. DNA methylation status of the SLC6A4 promoter region in hippocampal tissue was analyzed by bisulfite sequencing PCR (BSP).

Results

Compared with the blank group

the model group showed decreased sucrose preference

reduced total locomotor distance

and

prolonged latency to feeding (

0.05)

decreased serum 5-HT

and NE levels (

0.05)

downregulated hippocampal SLC6A4 and 5-HT1AR protein and mRNA expression (

0.05)

and increased CpG site methylation rate of the SLC6A4 promoter region (

0.05). Compared with the model group

the medication group

the 5-AZA group

and the EA group exhibited increased sucrose preference

increased total locomotor distance

shortened latency to feeding (

0.05)

elevated serum 5-HT

and NE levels (

0.05)

upregulated hippocampal SLC6A4 and 5-HT1AR protein and mRNA expression (

0.05)

and reduced CpG site methylation rate of the SLC6A4 promoter (

0.05). Compared with the medication group and the 5-AZA group

the EA group showed higher sucrose preference

greater total locomotor distance

shorter latency to feeding (

0.05)

and increased serum DA and NE levels (

0.05).

Conclusion

EA could improve depressive behaviors in depressed rat models. The underlying mechanism may involve inhibition of SLC6A4 hypermethylation in the hippocampus on the serotonergic system

upregulation of SLC6A4 and 5-HT1AR protein and mRNA expression

and elevation of monoamine neurotransmitters such as 5-HT.

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