Fang Hui, Wang Yan-Huan, Yang Lei, Che Yi-Hao, Liu Feng-Le, Liu Heng
Yunnan Provincial Key Laboratory of Entomological Biopharmaceutical R&D, National-Local Joint Engineering Research Center of Entomoceutics, Innovation Base for the Integrated Cultivation of Professional Master of Clinical Pharmacy and Clinical Pharmacist Standardized Training, College of Pharmacy, Dali University, Dali 671000, China; Department of Pharmacy, Dali Prefecture Traditional Chinese Medicine Hospital of Yunnan Province, Dali 671000, China.
Department of Pharmacy, NO.1 People's Hospital of Dali City, Dali 671000, China.
J Pharm Biomed Anal. 2026 Feb 15;269:117231. doi: 10.1016/j.jpba.2025.117231. Epub 2025 Nov 3.
Suanzaoren Tang (SZRT), a classic prescription from the Jingui Yaolue, is commonly used to treat insomnia associated with liver-blood deficiency. However, the mechanisms underlying SZRT's treatment of insomnia remain poorly understood. Using an integrated multi-omics approach, this research aimed to systematically explore the underlying mechanisms through which SZRT exerted its alleviating efficacy in p-chlorophenylalanine (PCPA)-induced insomnia. UPLC-ESI-MS/MS analysis was performed to identify the compounds in SZRT. Intraperitoneal administration of PCPA was utilized to develop an insomnia model in rats. Behavioral evaluation, ELISA, immunofluorescence, untargeted metabolomics and 16S rRNA sequencing were applied to assess how SZRT affected sleep quality, neurotransmitters, inflammatory markers, gut microbiota composition, and metabolic pathways. 30 compounds were identified in SZRT. PCPA treatment significantly prolonged sleep latency and altered neurochemical profiles, including decreased γ-aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT) levels, and increased glutamate (Glu), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and CYP2E1 expression. Immunofluorescence revealed reduced expression of glial fibrillary acidic protein (GFAP), ionized calcium-binding adapter molecule 1(Iba-1), 5-HTA, and 5-HTA receptors, indicating glial cell dysfunction and serotonergic dysregulation in PCPA group. SZRT intervention reversed these changes. Microbiota analysis showed that PCPA disrupted gut microbial composition, with decreased Prevotellaceae and Butyricicoccus abundance, both of which were restored by SZRT. Metabolomics identified key differential metabolites enriched in branched-chain amino acid metabolism, D-amino acid metabolism, α-linolenic acid metabolism, and arachidonic acid pathways. SZRT restored neurotransmitter homeostasis and reduced inflammation via microbiota-metabolite interactions. Suanzaoren Tang may improve insomnia by modulating the gut microbiota-metabolome-brain axis and glial cell activity, thereby restoring neurotransmitter balance, attenuating oxidative stress, and suppressing neuroinflammation. These findings provide experimental support for the clinical application of SZRT in insomnia treatment.
酸枣仁汤（SZRT）是《金匮要略》中的经典方剂，常用于治疗肝血不足所致的失眠。然而，酸枣仁汤治疗失眠的潜在机制仍不清楚。本研究采用综合多组学方法，系统探索酸枣仁汤对4-氯苯丙氨酸（PCPA）诱导的失眠发挥缓解作用的潜在机制。采用超高效液相色谱-电喷雾串联质谱（UPLC-ESI-MS/MS）分析鉴定酸枣仁汤中的化合物。通过腹腔注射PCPA建立大鼠失眠模型。采用行为学评价、酶联免疫吸附测定（ELISA）、免疫荧光、非靶向代谢组学和16S rRNA测序等方法，评估酸枣仁汤对睡眠质量、神经递质、炎症标志物、肠道微生物群组成和代谢途径的影响。在酸枣仁汤中鉴定出30种化合物。PCPA处理显著延长了睡眠潜伏期，并改变了神经化学特征，包括γ-氨基丁酸（GABA）和5-羟色胺（5-HT）水平降低，谷氨酸（Glu）、白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）和细胞色素P450 2E1（CYP2E1）表达增加。免疫荧光显示，胶质纤维酸性蛋白（GFAP）、离子钙结合衔接分子1（Iba-1）、5-羟色胺受体1A（5-HT1A）和5-羟色胺受体2A（5-HT2A）的表达降低，表明PCPA组存在神经胶质细胞功能障碍和5-羟色胺能调节异常。酸枣仁汤干预可逆转这些变化。微生物群分析表明，PCPA破坏了肠道微生物组成，普雷沃氏菌科和丁酸球菌丰度降低，而酸枣仁汤均可使其恢复。代谢组学鉴定出在支链氨基酸代谢、D-氨基酸代谢、α-亚麻酸代谢和花生四烯酸途径中富集的关键差异代谢物。酸枣仁汤通过微生物群-代谢物相互作用恢复神经递质稳态并减轻炎症。酸枣仁汤可能通过调节肠道微生物群-代谢组-脑轴和神经胶质细胞活性来改善失眠，从而恢复神经递质平衡、减轻氧化应激并抑制神经炎症。这些发现为酸枣仁汤在失眠治疗中的临床应用提供了实验依据。
