Effects of <italic style="font-style: italic">Hewei Anshe Formula</italic> （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models

WANG Shuo; WANG Changzhen; LI Zhihui; HUANG Tianke; WANG Liang; TIAN Chujiao; ZOU Tao; LIU Zihan; CHEN Qi; LI Shaodan

doi:10.13288/j.11-2166/r.2024.20.013

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Effects of Hewei Anshe Formula （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models

更新时间：2024-10-12

- Effects of Hewei Anshe Formula （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models
- Journal of Traditional Chinese Medicine Vol. 65, Issue 20, Pages: 2145-2151(2024)
- 作者机构：
  
  1.中国人民解放军总医院第六医学中心中医医学部，北京市海淀区阜成路6号，100048
  2.中国人民解放军医学院
  3.中国人民解放军军事科学院军事医学研究院
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.20.013
  CLC：
- Received：29 July 2024，
  
  Revised：2024-08-21，
  
  Published：17 October 2024
- 稿件说明：
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王硕,王长振,李志慧等.和胃安神方对失眠模型大鼠下丘脑生物钟基因CLOCK、BMAL1表达的影响[J].中医杂志,2024,65(20):2145-2151.

WANG Shuo,WANG Changzhen,LI Zhihui,et al.Effects of Hewei Anshe Formula （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models[J].Journal of Traditional Chinese Medicine,2024,65(20):2145-2151.
王硕,王长振,李志慧等.和胃安神方对失眠模型大鼠下丘脑生物钟基因CLOCK、BMAL1表达的影响[J].中医杂志,2024,65(20):2145-2151. DOI： 10.13288/j.11-2166/r.2024.20.013.

WANG Shuo,WANG Changzhen,LI Zhihui,et al.Effects of Hewei Anshe Formula （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models[J].Journal of Traditional Chinese Medicine,2024,65(20):2145-2151. DOI： 10.13288/j.11-2166/r.2024.20.013.

摘要

目的

探讨和胃安神方治疗失眠的可能作用机制。

方法

60只雄性SD大鼠随机分为正常组、模型组、艾司唑仑组与和胃安神方低、中、高剂量组。除正常组外其余各组大鼠采用腹腔注射对氯苯丙氨酸（PCPA）2天构建大鼠失眠模型。造模成功后，艾司唑仑组予以0.33 mg/（kg·d）艾司唑仑水溶液灌胃，和胃安神方低、中、高剂量组分别给予浓度为1、2、4 g/ml的和胃安神方10 ml/kg灌胃，正常组和模型组大鼠给予10 ml/kg的生理盐水灌胃，均每天1次，连续7天。实验期间观察大鼠一般情况，给药后每天测量大鼠体质量。最后一次给药后次日采用戊巴比妥钠协同实验观察各组大鼠睡眠情况，记录睡眠潜伏期及睡眠时长；采用免疫组化法和Western Blot法检测大鼠下丘脑时钟节律调节蛋白（CLOCK）、脑和肌肉芳香烃受体核转运样蛋白1（BMAL1）表达情况。

结果

模型组大鼠在各时间点体质量较正常组大鼠降低（

＜0.01）；与艾司唑仑组同时间比较，和胃安神方低剂量组大鼠在给药第5、6、7天体质量升高（

＜0.05）。与正常组比较，模型组大鼠睡眠时长缩短（

＜0.01）；与模型组比较，各给药组大鼠睡眠时长增加（

＜0.01）；和胃安神方高剂量组与艾司唑仑组睡眠时长比较差异无统计学意义（

＞0.05），而和胃安神方中、低剂量组睡眠时长较艾司唑仑组缩短（

＜0.01）。各组大鼠睡眠潜伏期比较差异均无统计学意义（

＞0.05）。免疫组化法和Western Blot法检测结果均显示，与正常组比较，模型组大鼠下丘脑中CLOCK、BMAL1表达显著减少（

＜0.01）；和胃安神方低、高剂量组大鼠下丘脑中CLOCK、BMAL1表达均较模型组增多（

＜0.05或

＜0.01）。

结论

和胃安神方可改善模型大鼠的失眠状况，其作用机制可能与上调下丘脑生物钟基因CLOCK、BMAL1蛋白表达有关。

Abstract

Objective

To investigate the possible mechanism of

Hewei Anshen Formula

（和胃安神方） in the treatment of insomnia.

Methods

Sixty male SD rats were randomly divided into the normal group， the model group， the eszop

iclone group and the low-， medium- and high-dose

Hewei Anshen Formula

groups. The insomnia model was constructed by intraperitoneal injection of p-chlorophenylalanine （PCPA） for 2 days in all groups except the normal group. After successful modelling， the eszopiclone group was given 0.33 mg/（kg·d） eszopiclone aqueous solution by gavage， the low-， medium- and high-dose

Hewei Anshen Formula

groups were given 10 ml/kg of

Hewei Anshen Formula

with a concentration of 1， 2 and 4 g/ml， respectively， and the rats in the normal group and the model group were given 10 ml/kg of saline by gavage， once a day for 7 consecutive days. The general condition of the rats was observed during the experiment， and the body mass of the rats was measured every day after medication administration. The following day after the last medication administration， pentobarbital sodium co-test was used to observe the sleep condition， and the sleep latency and sleep duration were recorded； immunohistochemistry and Western blot were used to detect the expression of hypothalamic clock rhythm regulating protein （CLOCK） and brain and muscle aromatic hydrocarbon receptor nuclear transporter-like protein 1 （BMAL1） in the rats.

Results

The body mass of rats in the model group was lower than that of rats in the normal group at all time points （

＜0.01）； compared with the same time in the eszopiclone group， the body mass of rats in the low-dose

Hewei Anshen Formula

group was elevated on the 5th， 6th and 7th days of medication administration （

＜0.05）. Compared with the normal group， the sleep duration of rats in the model group was shortened （

＜0.01）； compared with the model group， the sleep duration of rats in each dosage group increased （

＜0.01）， and the difference between the high-dose

Hewei Anshen Formula

group and the eszopiclone group showed no statistically significant （

＞0.05）， while the sleep duration of the low- and medium-dose

Hewei Anshen Formula

groups were shorterned than the eszopiclone group （

＜0.01）. The difference in sleep latency showed no statistically significant among each group （

＞0.05）. The results of both immunohistochemistry and Western blotting showed that the expression of CLOCK and BMAL1 in the hypothalamus of rats in the model group was significantly reduced compared with that in the normal group （

＜0.01）； the expression of CLOCK and BMAL1 in the hypothalamus of rats in the low- and high-dose

Hewei Anshen Formula

groups increased than that in the model group （

＜0.05 or

＜0.01）.

Conclusion

Hewei Anshen Formula

can improve insomnia in model rats， and its mechanism of action may be related to the up-regulation of the expression of the hypothalamic biological clock genes CLOCK and BMAL1 protein.

关键词

Keywords

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Effects of Hewei Anshe Formula （和胃安神方） on the CLOCK and BMAL1 Gene Expression of Hypothalamic Biological Clock in Insomnia Rat Models

DOI：10.13288/j.11-2166/r.2024.20.013

摘要

Abstract

关键词

Keywords

references