Insomnia is a common sleep disorder that severely affects patients' quality of life and physical and mental health. Abdominal massage, as a non-pharmacological therapy, has gradually attracted attention due to its definite therapeutic effect on insomnia and its safety with no significant side effects. However, the specific mechanism of abdominal massage in treating insomnia is still not clear.

We sought to scrutinise the distinctive hypothalamic protein expressions in p-chlorophenylalanine (PCPA) -provoked insomnia models in rats via proteomics, along with exploring the efficacy of abdominal massage in combating insomnia via behavioural analysis.

From March 2022 to May 2022, 18 healthy male Sprague-Dawley (SD) rats with a body weight of (225±5) g were selected. The SD rats were randomly divided into three groups: the blank group, the model group, and the treatment group, with six rats in each group. Insomnia was created via intraperitoneal administration of PCPA. The blank group was untreated while the model group received no intervention. The treatment group underwent abdominal massage, once a day for 12 minutes, for seven straight days. Observations were made on animal behaviour in each group. Proteomics technology identified differential protein expressions in the hypothalamus, followed by biological information analysis for Gene Ontology (GO) functional annotation and KEGG enrichment. Lastly, related differential proteins were validated by Western blotting.

The sodium pentobarbital righting reflex test was conducted 24 hours after modeling. The results showed that compared with the blank group, the model group and the treatment group had prolonged latency to sleep and shortened sleep duration (P<0.05), indicating that the modeling was successful in both groups. After 7 days of treatment, the behavioral results showed that compared with the blank group, the model group had decreased body weight, increased spontaneous paw-raising frequency, and prolonged time for rats to find the platform (P<0.05). Compared with the model group, the treatment group had increased body weight, reduced spontaneous paw-raising frequency, and shortened time for rats to find the platform (P<0.05). Proteomic analysis indicated that progerin-1 (Psen1) and creatine kinase (Ckm), linked to insomnia, were elevated post-modeling (P<0.05) and reduced post-treatment (P<0.05). γ-aminobutyric acid receptor subunit α-5 (Gabra5) and histone deacetylase 4 (Hdac4) were decreased post-modeling (P<0.05) and elevated post-treatment (P<0.05). GO and KEGG pathway enrichment analysis were found that these differentially expressed proteins are mainly involved in biological processes such as protein activation cascade, energy metabolism, inflammatory response and striated muscle tissue development. Neuroactive ligand-receptor interaction and hematopoietic cell lineage are the main signaling pathways of abdominal massage therapy.Western blotting validation of these four proteins confirmed three substances (Psen1, Ckm and Gabra5) displayed trends consistent with mass spectrometry.

The analysis of behavioral results and proteomics results indicates that abdominal massage can effectively improve insomnia symptoms in SD rats. It exerts its effects through the neuroactive ligand-receptor interaction and the hematopoietic cell lineage pathway. Psen1, Ckm, and Gabra5 are identified as potential key proteins. These findings provide a fundamental basis for the use of abdominal massage in the treatment of insomnia.

失眠是一种常见的睡眠障碍，严重影响患者的生活质量和身心健康。腹部推拿作为一种非药物疗法，因其在治疗失眠方面疗效确切且安全、无明显不良反应，逐渐受到关注，但其治疗失眠的具体机制尚不明确。

通过蛋白质组学分析，阐述腹部推拿治疗后对对氯苯丙氨酸（PCPA）失眠模型大鼠下丘脑蛋白的差异表达，结合行为学分析，探讨腹部推拿治疗失眠的作用机制。

2022年3—5月，取健康雄性Sprague-Dawley（SD）大鼠18只，体质量（225±5）g。将SD大鼠随机分为空白组、模型组和治疗组，每组6只。通过腹腔注射PCPA法建立大鼠失眠模型，空白组、模型组不做治疗，治疗组行腹部推拿，1次/d，12 min/次，连续7 d。观察记录各组大鼠行为状态，采用蛋白组学技术鉴定各组大鼠下丘脑差异蛋白表达情况，运用生物学信息进行差异蛋白基因本体论（GO）功能注释和KEGG富集分析，通过Western blotting法验证相关差异蛋白。

造模后24 h对大鼠进行戊巴比妥钠翻正反射实验，结果显示，模型组、治疗组与空白组相比，潜睡眠时间延长、睡眠持续时间缩短（P<0.05），提示造模成功。治疗7 d后，行为学结果显示，与空白组相比，模型组体质量下降、自发举爪次数增加、大鼠寻找平台的时间增加（P<0.05）；与模型组比较，治疗组体质量增加、自发举爪次数减少、大鼠寻找平台的时间缩短（P<0.05）。蛋白组学分析结果显示，与失眠关系密切的早老素-1（Psen1）和肌酸激酶（Ckm）造模后升高，治疗后降低（P<0.05）；γ-氨基丁酸受体亚基α-5（Gabra5）和组蛋白去乙酰化酶4（Hdac4）在造模后降低（P<0.05），治疗后升高（P<0.05）。GO和KEGG通路富集分析发现差异表达蛋白主要参与蛋白激活级联、能量代谢、炎症反应、横纹肌组织发育等生物过程，神经活性配体-受体相互作用、造血细胞谱系是腹部推拿治疗作用的主要信号通路。对这以上4种蛋白进行Western blotting法验证，其中3种物质（Psen1、Ckm和Gabra5）显示出与质谱显示的趋势一致。

经行为学结果及蛋白组学结果分析表明，腹部推拿可有效改善SD大鼠失眠症状，通过神经活性配体-受体相互作用、造血细胞谱系通路发挥作用，Psen1、Ckm、Gabra5为可能的关键蛋白，为腹部推拿治疗失眠提供了基础性依据。