Effect of <italic style="font-style: italic">Tongdu Tiaoshen</italic> acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway

PAN Wen; ZHANG Daozong; JIAO Shiyi; ZOU Ling; ZHANG Xiaomeng; QIAN Tingting; GAO Zhi; CHU Yifan; OU Kaiyan; LIU Feiyang; FU Xin; SUN Peiyang

doi:10.13703/j.0255-2930.20250629-k0002

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Effect of Tongdu Tiaoshen acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway

Study on Mechanism | 更新时间：2026-07-09

- Effect of Tongdu Tiaoshen acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway
- Chinese Acupuncture & Moxibustion Vol. 46, Issue 7, Pages: 1125-1135(2026)
- 作者机构：
  
  安徽中医药大学研究生院，合肥　230012
  安徽中医药大学第二附属医院，合肥　230061
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20250629-k0002
  CLC：
- Received：29 June 2025，
  
  Online First：27 April 2026，
  
  Published：12 July 2026
- 稿件说明：
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PAN Wen, ZHANG Daozong, JIAO Shiyi, et al. Effect of Tongdu Tiaoshen acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway[J]. Chinese Acupuncture & Moxibustion, 2026, 46(7): 1125-1135. DOI: 10.13703/j.0255-2930.20250629-k0002.
DOI：

PAN Wen, ZHANG Daozong, JIAO Shiyi, et al. Effect of Tongdu Tiaoshen acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway[J]. Chinese Acupuncture & Moxibustion, 2026, 46(7): 1125-1135. DOI: 10.13703/j.0255-2930.20250629-k0002. DOI：

摘要

目的：

观察“通督调神”针刺对慢性不可预知温和刺激（CUMS）大鼠抑郁样行为及微小RNA-26a-5p（miR-26a-5p）/Zeste基因增强子同源物2（EZH2）/组蛋白H3第27位赖氨酸三甲基化（H3K27me3）蛋白和炎症因子表达的影响，探讨其抗抑郁机制。

方法：

将32只SPF级雄性SD大鼠随机分为正常组（8只）、造模组（24只），造模组采用CUMS法建立抑郁模型，造模成功后随机分为模型组、针刺组、氟西汀组，每组8只。针刺组行“通督调神”针刺，穴取“百会”“神庭”“水沟”“大椎”，每日1次，干预6 d后休息1 d，共干预24次。氟西汀组灌胃氟西汀溶液（2.1 mg/kg），其余3组大鼠灌胃0.9%氯化钠溶液（10 mL/kg），连续28 d。观察大鼠蔗糖偏嗜度及旷场实验结果；HE和Nissl染色法观察海马神经元形态和尼氏小体；Western blot法检测海马EZH2、H3K27me3蛋白表达；RT-qPCR法检测海马miR-26a-5p、EZH2 mRNA表达；ChIP-qPCR法检测H3K27me3在白细胞介素（IL）-4、IL-6、肿瘤坏死因子-α（TNF-α）基因启动子区的富集水平；ELISA法检测海马及血清IL-6、TNF-α、IL-1β、IL-4含量。

结果：

与正常组比较，模型组蔗糖偏嗜度降低（

0.01），旷场实验运动总距离、直立次数及清洁行为减少（

0.01）；海马神经元部分溶解、排列不齐、核固缩，尼氏小体减少（

0.01），分布不均；海马EZH2 mRNA、EZH2及H3K27me3蛋白表达升高（

0.01），miR-26a-5p mRNA表达降低（

0.01）；IL-6、TNF-α基因启动子区H3K27me3富集水平升高（

0.01），IL-4基因启动子区H3K27me3富集水平降低（

0.01）；海马及血清IL-6、TNF-α、IL-1β含量升高（

0.01），IL-4含量降低（

0.01）。与模型组比较，针刺组和氟西汀组蔗糖偏嗜度增高（

0.01），旷场实验运动总距离、直立次数、清洁行为增加（

0.01）；神经元溶解减少、排列相对整齐、核固缩减少，尼氏小体增多（

0.01），分布较为均匀；海马EZH2 mRNA、EZH2及H3K27me3蛋白表达降低（

0.01），miR-26a-5p mRNA表达升高（

0.01），miR-26a-5p mRNA与EZH2 mRNA表达水平呈负相关（

0.001）；IL-6、TNF-α基因启动子区H3K27me3富集水平降低（

0.01），IL-4基因启动子区H3K27me3富集水平升高（

0.01，

0.05）；海马及血清IL-6、TNF-α、IL-1β含量降低（

0.01），IL-4含量升高（

0.01）。针刺组和氟西汀组各指标比较，差异无统计学意义（

0.05）。

结论：

“通督调神”针刺可改善CUMS大鼠抑郁样行为，其机制可能与调节miR-26a-5p/EZH2/H3K27me3通路，减轻中枢神经系统炎症有关。

Abstract

Objective

To observe the effects of

Tongdu Tiaoshen

(unblocking the circulation of the governor vessel and regulating the mind) acupuncture on depressive-like behavior and the expression of microRNA-26a-5p (miR-26a-5p)/ enhancer of zeste homolog 2 (EZH2)/trimethylation of lysine 27 on histone H3 (H3K27me3) proteins and inflammatory factors in chronic unpredictable mild stress (CUMS) rats

and to explore its antidepressant mechanism.

Methods

Thirty-two SPF-grade male SD rats were randomly divided into a normal gro

up (8 rats) and a modeling group (24 rats). The modeling group was subjected to CUMS to establish depression model. After successful modeling

the rats were randomized into a model group

an acupuncture group and a fluoxetine group

with 8 rats in each group. In the acupuncture group

Tongdu Tiaoshen

acupuncture was operated at "Baihui" (GV20)

"Shenting" (GV24)

"Shuigou" (GV26) and "Dazhui" (GV14)

once daily for 6 consecutive days followed by one day of interval; and the completion of intervention was composed of 24 sessions of acupuncture. The gavage with fluoxetine solution (2.1 mg/kg) was administered in the fluoxetine group; and with 0.9% sodium chloride solution (10 mL/kg) in the other 3 groups

continuously for 28 days. Sucrose preference and the results of the open field test were observed. The hippocampal neuron morphology and Nissl bodies were examined using HE and Nissl staining. The protein expression of hippocampal EZH2 and H3K27me3 was detected using Western blot

and the mRNA expression of hippocampal miR-26a-5p and EZH2 was detected with RT-qPCR. ChIP-qPCR was applied to detect the enrichment levels of H3K27me3 in the promoter regions of interleukin-4 (IL-4)

interleukin-6 (IL-6)

and tumor necrosis factor-α (TNF-α). ELISA was provided to measure the contents of IL-6

TNF-α

interleukin-1β (IL-1β)

and IL-4 in the hippocampus and serum.

Results

Compared with the normal group

the model group showed the decrease in sucrose preference (

0.01)

the total distance

the numbers of uprighting and cleaning behavior in the open field test (

0.01); hippocampal neurons exhibited partial cell dissolution

irregular arrangement

nuclear pyknosis

reduced Nissl bodies (

0.01)

and uneven distribution. The mRNA expression of hippocampal EZH2

and the protein expression of hippocampal EZH2 and H3K27me3 increased (

0.01)

while the mRNA expression of miR-26a-5p decreased (

0.01). The enrichment levels of H3K27me3 in the promoter regions of IL-6 and TNF-α were elevated (

0.01)

whereas that in IL-4 was reduced (

0.01). The levels of IL-6

TNF-α and IL-1β in the hippocampus and serum increased (

0.01)

and the levels of IL-4 decreased (

0.01). Compared with the model group

the acupuncture group and the fluoxetine group showed the increase in sucrose preference (

0.01)

the total distance

the numbers of uprighting and cleaning behavior in the open field test (

0.01); hippocampal neurons exhibited the reduced cell dissolution

regular arrangement

declined nuclear pyknosis

higher Nissl bodies (

0.01)

and even distribution. The mRNA expression of hippocampal EZH2

the protein expression of hippocampal EZH2 and H3K27me3 decreased (

0.01)

and the mRNA expression of miR-26a-5p increased (

0.01); and the mRNA expression of miR-26a-5p was negatively correlated with that of EZH2 (

0.001). The enrichment levels of H3K27me3 in the promoter regions of IL-6 and TNF-α were lower (

0.01)

and that of IL-4 was higher (

0.01

0.05). The levels of IL-6

TNF-α and IL-1β in the hippocampus and serum decreased (

0.01)

and the levels of IL-4 increased (

0.01). The differences of the above indexes were not statistically significant between the acupuncture group and fluoxetine group (

0.05).

Conclusion

Tongdu Tiaoshen

acupuncture can ameliorate depression-like behavior in CUMS rats

with the mechanism potentially related to regulating the miR-26a-5p/EZH2/ H3K27me3 pathway and reducing central neuroinflammation.

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⁰

Effect of Tongdu Tiaoshen acupuncture on neuroinflammation in rats with CUMS based on the miR-26a-5p/EZH2/H3K27me3 pathway

DOI：10.13703/j.0255-2930.20250629-k0002

摘要

Abstract

关键词

Keywords

references