Table2_Evaluation of the pharmacological effects and exploration of the mechanism of traditional Chinese medicine preparation Ciwujia tablets in treating insomnia based on ethology, energy metabolism, and urine metabolomic approaches.xlsx

Ciwujia Tablets (CWT) are produced by concentrating and drying the extract solution of the dried rhizome of Eleutherococcus senticosus (Rupr. & Maxim.) Maxim [Araliaceae; E. senticosus radix et rhizoma]. Besides, CWT is included in the 2020 edition of Chinese Pharmacopoeia and is widely used in the treatment of insomnia. It mainly contains eleutheroside B, eleutheroside E, isofraxidin, eleutheroside C, ciwujiatone, and chlorogenic acid, as well as other chemical components. Although the clinical efficacy of CWT in treating insomnia has been confirmed, its functions and pharmacological effects have not been systematically evaluated and its mechanism of action in the treatment of insomnia remains unclear. Therefore, in this study, behavioral, energy metabolism, and metabonomics methods were applied to systematically evaluate the effect of CWT on insomnia. Additionally, urine metabonomics based on UPLC-Q-TOF-MS/MS were utilized to identify potential endogenous biomarkers of insomnia, detect the various changes before and after CWT treatment, explore the metabolic pathway and potential target of CWT, and reveal its pharmacological mechanism. Results revealed that CWT increased inhibitory neurotransmitter (5-HT and GABA) content and reduced the content of excitatory neurotransmitters (DA and NE). Moreover, CWT enhanced autonomous behavioral activity, stabilized emotions, and promoted the return of daily basic metabolic indexes of insomniac rats to normal levels. The urine metabolomics experiment identified 28 potential endogenous biomarkers, such as allysine, 7,8-dihydroneopterin, 5-phosphonooxy-L-lysine, and N-acetylserotonin. After CWT treatment, the content of 22 biomarkers returned to normal levels. The representative markers included N-acetylserotonin, serotonin, N-methyltryptamine, and 6-hydroxymelatonin. Additionally, the metabolic pathways in rats were significantly reversed, such as tryptophan metabolism, folate biosynthesis, phenylalanine metabolism, and tyrosine metabolism. Ultimately, it is concluded that CWT regulated tryptophan metabolism, folate biosynthesis, phenylalanine metabolism, and other metabolic levels in the body. This drug has been confirmed to be effective in the treatment of insomnia by regulating the content of serotonin, 6-hydroxymelatonin, N-acetylserotonin, and N-methyltryptamine to a stable and normal level in tryptophan metabolism.

刺五加片（Ciwujia Tablets, CWT）系由刺五加[Eleutherococcus senticosus (Rupr. & Maxim.) Maxim，五加科：刺五加根及根茎]的干燥根茎提取液经浓缩干燥制得。此外，CWT收载于《中华人民共和国药典》2020年版，临床广泛用于失眠症的治疗。其主要成分为刺五加苷B、刺五加苷E、异秦皮啶、刺五加苷C、刺五加酮、绿原酸及其他多种化学成分。尽管刺五加片治疗失眠的临床疗效已得到证实，但其功能与药理作用尚未得到系统评价，治疗失眠的作用机制仍不明确。为此，本研究采用行为学、能量代谢及代谢组学方法，系统评价刺五加片对失眠症的干预效果。此外，本研究基于超高效液相色谱-串联四极杆飞行时间质谱（UPLC-Q-TOF-MS/MS）的尿液代谢组学技术，鉴定失眠潜在内源性生物标志物，检测刺五加片治疗前后的代谢变化，探究其代谢通路与潜在靶点，揭示其药理作用机制。研究结果显示，刺五加片可升高抑制性神经递质5-羟色胺（5-HT）与γ-氨基丁酸（GABA）的含量，降低兴奋性神经递质多巴胺（DA）与去甲肾上腺素（NE）的水平。此外，刺五加片可增强失眠模型大鼠的自主行为活动、稳定其情绪，并使其日常基础代谢指标恢复至正常水平。尿液代谢组学实验共鉴定出28种潜在内源性生物标志物，包括醛赖氨酸、7,8-二氢新蝶呤、5-磷酸氧基-L-赖氨酸、N-乙酰血清素等。经刺五加片治疗后，其中22种生物标志物的含量恢复至正常水平，代表性标志物包括N-乙酰血清素、血清素、N-甲基色胺与6-羟基褪黑素。此外，失眠模型大鼠的代谢通路得到显著逆转，如色氨酸代谢、叶酸生物合成、苯丙氨酸代谢及酪氨酸代谢通路。最终研究结论表明，刺五加片可调节机体内色氨酸代谢、叶酸生物合成、苯丙氨酸代谢等代谢水平。该药物通过调节色氨酸代谢通路中5-羟色胺、6-羟基褪黑素、N-乙酰血清素与N-甲基色胺的含量至稳定正常水平，从而证实其治疗失眠的有效性。