Huajuan Lei ^1,2 Jian Liu ^2,3* Juan Deng ^2,4* Pan Zou ^5* Zixiang Zou ^2,6* Ziou Li ^2* Honghui Li ^7* Zhoujin Tan ^2,8* Lin Luo ^8*

• ¹ Department of anesthesiology, the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Traditional Chinese Medicine of ChangSha medical college, The First Hospital of Hunan University of Chinese Medicine, Changsha, China
• ² Hunan University of Chinese Medicine, Changsha, Anhui Province, China
• ³ Department of Innovation Experiments Center,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine, The First Hospital of Hunan University of Chinese Medicine, Changsha, China
• ⁴ The third hospital of changsha,Department of anesthesiology,, The First Hospital of Hunan University of Chinese Medicine, Changsha, China
• ⁵ department of radiology,the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine, The First Hospital of Hunan University of Chinese Medicine, Changsha, China
• ⁶ the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine,Department of gynaecology, The First Hospital of Hunan University of Chinese Medicine, Changsha, China
• ⁷ Department of orthopedics, Hunan University of Chinese Medicine, Changsha, Anhui Province, China
• ⁸ The First Hospital of Hunan University of Chinese Medicine, Changsha, China

practice and is characterized by depression and autonomic nervous symptoms, including hot flashes, palpitation, and night sweating. In addition, the comorbidity of Menopause depression has long been an integral component of the estradiol ( E2 ) shortage. Previous studies have suggested that the mechanisms underlying this comorbidity involved overlap of endocrine and cerebellar networks. Emerging evidence has shown that the endocrine-brain-gut-micobiota axis plays a key role in the regulation of affective disorders. Thus, we hypothesized that the YYNS may involved in the menopause depression alleviation through endocrine-brain-gut-micobiota axis. To verify this, we constructed a bilateral ovariectomy (OVX) mouse model to simulate menopausal-related depression. Subsequently, behavioral tests including the Open Field Test (OFT) and the Forced Swimming Test (FST) were conducted to examine the depression state post-OVX. With YYNS or E2 intervention, enzyme-linked immunosorbent assay ( ELISA ) was used to determine the serum sex hormones level. 16S rRNA gene sequencing and liquid chromatography-mass spectrometry ( LC-MS ) were used to analyze the microbiome of the colon samples collected from mice in the sham surgery group ( CSH ), the OVX model group ( CMD ), the OVX with E2 hormone intervention group ( CHM ), and the OVX with YYNS intervention group ( CYYNS ). One week after OVX, CMD, CHM and CYYNS showed depression in OFT, FST 3 week post-OVX, CHM and CYYNS showed a depression relieved notably, OVX shaped the OTUs shrinkage, OTUs raised in the Sham, CHM and CYYNS group. CMD group showed abundance of Actinobiota decreased, but of Bacteriodia increased. The relative abundance of genus varied in each group.Moreover, functional correlation of changes in Sex hormone and gut microbes between different groups showed that the PRL level was negatively correlated with Odoribacter. T level was positively correlated with Lachnospiraceae NK4A136 group and Odoribacter abundance ( P< 0.05 ). Our results not only offer novel insights into the sex hormones and depression with OVX mice, but also build an important basis for E2 or YYNS therapeutic efficacy on PDD which provide for future research on this etiology through endocrine-brain-gut-microbiota network.