Zhang Chunge, Wu Qi, Tao Xiang, Yan Zhaowei, Han Qiang, Yao Xin, Chai Yuying, Chen Lin, Mao Yeqin, Cheng Zongqi
Department of Pharmacy, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
Department of Pharmacy, the First Affiliated Hospital of Soochow University, Suzhou 215006, China.
J Pharm Biomed Anal. 2025 Sep 15;263:116949. doi: 10.1016/j.jpba.2025.116949. Epub 2025 May 5.
This study aims to elucidate the pathways through which Yiyin Anshen Granule (YA) exerts sedative-hypnotic effects in a mouse model of sleep deprivation. DL-4-chlorophenylalanine(PCPA)-treated mice received intragastric administration of YA and pentobarbital sodium-induced sleep tests were conducted on days 7, 14, and 29. The levels of key neurotransmitters, cytokines and receptor protein associated with insomnia were measured using enzyme-linked immunosorbent assay (ELISA) and Western blot. Additionally, 16S ribosomal DNA (rDNA) sequencing was performed to assess the impact of YA on gut microbiota, focusing on species abundance and diversity. YA significantly shortened sleep latency (P < 0.01) and prolonged sleep duration (P < 0.01) in sleep-deprived mice, effectively improving circadian rhythm disturbances compared to the model group (MOD). Biochemical analysis revealed that YA restored abnormal neurotransmitter levels in brain tissue, increasing 5-hydroxytryptamine (5-HT), γ-aminobutyric acid (GABA), and γ-aminobutyric acid type A receptor α-1 subunit (GABAARα1) expression (P < 0.01) and reducing the glutamate (Glu)/GABA ratio (P < 0.01). Additionally, the levels of B-cell lymphoma 2 (BCL-2) and interleukin-6 (IL-6) expression were significantly decreased (P < 0.05, 0.01), while interleukin-1 beta (IL-1β) expression increased (P < 0.01). YA treatment also significantly increased gut microbiota abundance and diversity, with microbiome profiles in the YA group being closer than those of diazepam group (DZP) to the control group (CON). Notably, YA reversed the dysbiosis of high-abundance gut microbiota species associated with insomnia at both the family and genus levels (P < 0.05, 0.01). The results of the present study indicated that YA alleviates insomnia symptoms by regulating neurotransmitter and inflammatory cytokines levels, and restoring gut microbia balance. These mechanisms collectively contribute to shortening sleep latency, prolonging sleep duration, and improving sleep quality in a mouse model of insomnia.
本研究旨在阐明益阴安神颗粒（YA）在睡眠剥夺小鼠模型中发挥镇静催眠作用的途径。用DL-4-氯苯丙氨酸（PCPA）处理的小鼠接受YA灌胃给药，并在第7、14和29天进行戊巴比妥钠诱导的睡眠试验。使用酶联免疫吸附测定（ELISA）和蛋白质印迹法测量与失眠相关的关键神经递质、细胞因子和受体蛋白水平。此外，进行16S核糖体DNA（rDNA）测序以评估YA对肠道微生物群的影响，重点关注物种丰度和多样性。与模型组（MOD）相比，YA显著缩短了睡眠剥夺小鼠的睡眠潜伏期（P<0.01）并延长了睡眠时间（P<0.01），有效改善了昼夜节律紊乱。生化分析表明，YA恢复了脑组织中异常的神经递质水平，增加了5-羟色胺（5-HT）、γ-氨基丁酸（GABA）和γ-氨基丁酸A型受体α-1亚基（GABAARα1）的表达（P<0.01），并降低了谷氨酸（Glu）/GABA比值（P<0.01）。此外，B细胞淋巴瘤2（BCL-2）和白细胞介素-6（IL-6）的表达水平显著降低（P<0.05，0.01），而白细胞介素-1β（IL-1β）的表达增加（P<0.01）。YA治疗还显著增加了肠道微生物群的丰度和多样性，YA组的微生物群谱比地西泮组（DZP）更接近对照组（CON）。值得注意的是，YA在家族和属水平上逆转了与失眠相关的高丰度肠道微生物物种的生态失调（P<0.05，0.01）。本研究结果表明，YA通过调节神经递质和炎性细胞因子水平以及恢复肠道微生物平衡来减轻失眠症状。这些机制共同有助于缩短失眠小鼠模型的睡眠潜伏期、延长睡眠时间并改善睡眠质量。
