He ' s Yangchao Formula Ameliorates Premature Ovarian Insuf-ficiency via Remodeling Gut Microbiota to Promote Granulosa Cell Glycolysis

• Published in: Zhejiang da xue xue bao. Journal of Zhejiang University. Medical sciences. Yi xue ban p. 1
• Main Authors: LIN, Fangxuan, LIU, Qing, ZHAO, Ying, CHEN, Yun, WANG, Ruye, MIAO, Chenyun, ZHANG, Qin
• Format: Journal Article
• Language: English
• Published: China 04.07.2025
• Subjects: Glycolysis; Mice; Intestinal flora; Premature ovarian insufficiency; He’s Yangchao recipe
• ISSN: 1008-9292
• DOI: 10.3724/zdxbyxb-2024-0604

To investigate the molecular mechanism by which He's Yangchao Formula improves ovarian function in premature ovarian insufficiency (POI) mice through intestinal flora modulation. Forty female ICR mice (aged 6-8 weeks) were intraperitoneally injected with cyclophosphamide (150 mg/kg) to establish a POI model, while 10 untreated mice served as the blank control. Successfully modeled mice were randomly divided into four groups ( =10/group): low-dose He's Yangchao Formula (6 g crude herb/kg), high-dose He's Yangchao Formula (25 g crude herb/kg), positive control (estradiol), and model control (distilled water). Treatments were admin-istered daily by gavage for 6 weeks. Vaginal exfoliated cells were stained with Wright-Giemsa solution to monitor estrous cycles. Serum estradiol and follicle-stimulating hormone (FSH) levels were measured by ELISA. Ovarian FSH receptor (FSHR) expression was assessed by immunohistochemistry. Glycolysis-related proteins pyruvate kinase M2 (PKM2) and glucose transporter 4 (GLUT4) were analyzed by Western blotting and immuno-fluorescence. Fecal samples from blank control, model control, and high-dose groups underwent metagenomic sequencing to evaluate intestinal microbiota diversity and com-position. He's Yangchao Formula restored estrous cyclicity, increased serum estradiol ( <0.05), decreased serum FSH ( <0.05), and upregulated FSHR expression in granulosa cells ( <0.05). Metagenomic analysis revealed significant structural differences in intestinal flora among blank control, model control, and high-dose groups ( <0.01). The high-dose group showed reduced abundance of conditional pathogens (e.g., , , , , ) compared to the model control ( <0.05). Functional enrichment analysis indicated involvement of glycolysis-related pathways. Concordantly, PKM2 and GLUT4 expression was downregulated in the model control but upregulated in He's Yangchao Formula groups ( <0.05). He's Yangchao Formula ameliorates POI in mice by remodeling intestinal flora structure, enhancing glycolytic activity, improving ovarian sex hormone secretion, increasing granulosa cell FSHR expression, and restoring estrous cyclicity.