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Introduction: Premenstrual dysphoric disorder (PMDD) is a cyclical mooddisorder that severely affects the daily life of women of reproductive age.Most of the medications being used clinically have limitations such as lowefficacy, side effects, and high cost, so there is an urgent need to discoversafer and more effective medications. Rutin is a natural flavonol glycoside withvarious pharmacological properties including antidepressant. The study of theefficacy and mechanism of action of rutin in PMDD-depressed subtype modelrats plays an important role in the discovery of new drugs for thetreatment of PMDD.Methods: Binding of rutin to gamma-aminobutyric acid type A receptors (GABAAreceptors) was probed using molecular docking, microscale thermophoresis,radioactive receptor ligand binding assay and cell membrane clamp experiment.Behavioral tests in mice were performed to screen the optimal dose of rutin.Behavioral tests were performed to evaluate the effects of rutin on depressedmood, memory impairment, and social impairment in PMDD-depressed subtypemodel rats. HE staining and Golgi staining were performed to observe theneuronal damage in rat hippocampus. UHPLC-MS/MS targeted metabolomicswas performed to detect the changes of neurotransmitter content in rathippocampus. PCR array to detect the effect of rutin on mRNA expression ofGABAA receptor partial subunits in rat hippocampus.Results: The docking score of rutin with the GABAA receptor benzodiazepine site was −11.442 and the gliding score was −11.470. The Kd of rutin with the GABAA receptor (α1β2γ2) was 1.17 ± 0.89 μM. Rutin competed with [H3 ]-flunitrazepam for the GABAA receptor benzodiazepine site and inhibited the inward flow of chloride ions (P < 0.05). In PMDD-depressed subtype rats, rutin alleviated depressed mood, memory impairment and social impairment, ameliorated hippocampal neuronal damage and reduces gamma-aminobutyric acid (GABA) and acetylcholine (ACh) levels (P < 0.05). Moreover, we found that rutin did not affect the relative mRNA expression of GABAA receptor subunits in rat hippocampus.Discussion: Overall, rutin alleviated depressed mood, memory impairment andsocial impairment in PMDD-depressed subtype rats, which may be related tobinding to GABAA receptor benzodiazepine sites, inhibiting chloride ions inwardflow, ameliorating hippocampal neuronal damage and reducing GABA and AChlevels. The results of this study provide an experimental basis and scientificevidence for the development of new drugs for the treatment of PMDD.

引言：经前期烦躁障碍（Premenstrual dysphoric disorder, PMDD）是一种周期性情绪障碍，严重影响育龄期女性的日常生活。当前临床使用的多数药物存在疗效低下、不良反应明显且成本高昂等局限性，因此亟需开发更安全有效的治疗药物。芦丁是一种天然黄酮醇苷，具有包括抗抑郁活性在内的多种药理特性。针对芦丁在经前期烦躁障碍抑郁亚型模型大鼠中的疗效及作用机制开展研究，可为开发治疗PMDD的新型药物提供重要依据。
方法：采用分子对接、微量热泳动技术、放射性受体配体结合实验及细胞膜片钳实验，探究芦丁与A型γ-氨基丁酸受体（gamma-aminobutyric acid type A receptors, GABAA受体）的结合特性。通过小鼠行为学实验筛选芦丁的最优给药剂量；继而通过行为学实验评估芦丁对经前期烦躁障碍抑郁亚型模型大鼠抑郁样情绪、记忆损伤及社交功能受损的改善作用。采用苏木精-伊红染色（hematoxylin-eosin staining, HE染色）与高尔基染色（Golgi staining）观察大鼠海马神经元损伤情况；通过超高效液相色谱-串联质谱（ultra high performance liquid chromatography-tandem mass spectrometry, UHPLC-MS/MS）靶向代谢组学检测大鼠海马组织内神经递质含量的变化；利用PCR基因表达阵列（PCR array）检测芦丁对大鼠海马组织内GABAA受体部分亚基mRNA表达水平的影响。
结果：芦丁与GABAA受体苯二氮䓬结合位点的对接得分为-11.442，滑行得分为-11.470。芦丁与GABAA受体（α1β2γ2亚型）的解离常数Kd为1.17±0.89 μM。芦丁可与[³H]-氟硝西泮竞争结合GABAA受体苯二氮䓬位点，并抑制氯离子内流（P<0.05）。在经前期烦躁障碍抑郁亚型模型大鼠中，芦丁可改善抑郁样情绪、缓解记忆损伤与社交功能受损，减轻海马神经元损伤，并降低γ-氨基丁酸（GABA）及乙酰胆碱（ACh）的含量（P<0.05）。此外，本研究发现芦丁对大鼠海马组织内GABAA受体亚基的相对mRNA表达水平无显著影响。
讨论：综上，芦丁可改善经前期烦躁障碍抑郁亚型模型大鼠的抑郁样情绪、记忆损伤及社交功能受损，其作用机制可能与结合GABAA受体苯二氮䓬位点、抑制氯离子内流、减轻海马神经元损伤及降低GABA与ACh含量相关。本研究结果为开发治疗PMDD的新型药物提供了实验基础与科学依据。