Xiangjun Wang ^1* Xiaowen Xia ¹ Xianliang Song ^2* Yi Zhou ³ Mingyu Ma ² Yashuang Ren ^2* Xitai Chen ^2* Zenghui Xia ^1* Yinghui Guo ^4* Chunhong Song ^1*

• ¹ Laboratory Animal Center, Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China
• ² School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China
• ³ Department of Pharmacy, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, China
• ⁴ College of Chinese Medicine, Shandong University of Traditional Chinese Medicine, Jinan, Shandong Province, China

Introduction: Premenstrual dysphoric disorder (PMDD) is a cyclical mood disorder that severely affects the daily life of women of reproductive age. Most of the medications being used clinically have limitations such as low efficacy, side effects, and high cost, so there is an urgent need to discover safer and more effective medications. Rutin is a natural flavonol glycoside with various pharmacological properties including antidepressant. The study of the efficacy and mechanism of action of rutin in PMDD-depressed subtype model rats plays an important role in the discovery of new drugs for the treatment of PMDD.Methods: Binding of rutin to GABAA receptors 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 optimal dose of rutin. Behavioral tests were performed to evaluate the effects of rutin on depressed mood, memory impairment, and social impairment in PMDD-depressed subtype model rats. HE and Golgi staining were performed to observe the neuronal damage. UHPLC-MS/MS targeted metabolomics was performed to detect the changes of neurotransmitter content. PCR array to detect the effect of rutin on mRNA expression of GABAA receptor subunits.Results: The docking score of rutin with the GABAA receptor benzodiazepine site was -11.442 and gliding score was -11.470. The Kd of rutin with the GABAA receptor was 1.17 ± 0.89 μM. Rutin competed with [H 3 ]-flunitrazepam for the GABAA receptor benzodiazepine site and inhibited inward flow of chloride (P < 0.05). In PMDD-depressed subtype rats, rutin alleviated depressed mood, memory impairment and social impairment, ameliorated hippocampal neuronal damage and reduces GABA and acetylcholine levels (P < 0.05). Moreover, we found that rutin did not affect the relative mRNA expression of GABAA receptor subunits in hippocampus. Discussion: Overall, rutin alleviated depressed mood, memory impairment and social impairment in PMDD-depressed subtype rats, which may be related to binding to GABAA receptor benzodiazepine sites, inhibiting chloride inward flow, ameliorating hippocampal neuronal damage and reducing GABA and ACh levels. The results of this study provide an experimental basis and scientific evidence for the development of new drugs for the treatment of PMDD.