Mechanistic study on the holistic actions of Hong-Hua-Xiao-Yao Tablet in alleviating premenstrual syndrome in rats

  • Phytomedicine. 2026 Jun:155:158161. doi: 10.1016/j.phymed.2026.158161.
Wenjing Wei  1 ,  Mingxiao Li  2 ,  Kedian Chen  3 ,  Xiaofang He  4 ,  Shuhui Tang  4 ,  Huaijue Deng  4 ,  Weize Zhu  2 ,  Yuanyuan Li  4 ,  Lili Sheng  4 ,  Guangbo Ge  5 ,  Jianbo Wan  6 ,  Na Lin  3 ,  Yanqiong Zhang  7 ,  Ningning Zheng  8 ,  Houkai Li  9
Affiliations
  • 1. Institute of Pediatric Infection, Immunity, and Critical Care Medicine, Shanghai Children's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200062, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 2. Institute of Pediatric Infection, Immunity, and Critical Care Medicine, Shanghai Children's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200062, China.
  • 3. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
  • 4. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 5. Shanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
  • 6. The State Key Laboratory of Mechanism and Quality of Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China.
  • 7. Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China. Electronic address: [email protected].
  • 8. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. Electronic address: [email protected].
  • 9. Institute of Pediatric Infection, Immunity, and Critical Care Medicine, Shanghai Children's Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200062, China; School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China; Guangdong Provincial Key Laboratory of Chinese Medicine Ingredients and Gut Microbiomics, School of Pharmacy, Shenzhen University Medical School, Shenzhen 518060, China. Electronic address: [email protected].
PMID: 41990531 DOI: 10.1016/j.phymed.2026.158161
Abstract

Background: Premenstrual syndrome (PMS) is a cyclical neuroendocrine disorder that adversely affects the physical and mental health of women globally. Hong-Hua-Xiao-Yao Tablet (HHXYT) is a marketed Chinese patent medicine, with well-established therapeutic effect on PMS in clinic. However, its underlying mechanisms are poorly understood due to the extreme complexity of both PMS and HHXYT itself.

Purpose: To elucidate the systemic mechanisms of action of HHXYT ameliorates PMS.

Methods: An integrated strategy combining serum metabolomics and tissue-specific transcriptomics was applied to identify key metabolic pathways affected by HHXYT. Additionally, 16S rRNA gene Sequencing was employed to evaluate gut microbiota composition. In vitro assays were performed to assess anti-neuroinflammatory effects in PMS-related cells. The bioactivity of blood-absorbed compounds was further verified. The role of gut microbiota was further investigated using ex vivo cultures and pseudo germ-free models.

Results: HHXYT affected the serum metabolite profiles and target organ transcriptional profiles in the glucose 6-phosphate metabolism, sphingolipid metabolism, glycerophospholipid metabolism, which in turn improved the systemic symptoms of PMS. Moreover, HHXYT-containing serum specifically inhibited neuroinflammation in microglia, without affecting Other PMS-associated target cells. Subsequently, we identified 62 blood-absorbed components, primarily derived from Glycyrrhiza uralensis (GC, 36 compounds). Among these, four GC-derived compounds-glycyrrhizic acid, 18β-glycyrrhetinic acid, liquiritin, and liquiritigenin significantly suppressed neuroinflammation in microglia. Interestingly, the lack of detected blood-absorbed components from the sovereign herb Bupleurum marginatum implies that its therapeutic benefits may occur via an indirect mechanism involving gut microbiota metabolism. Using ex vivo cultures and pseudo germ-free models, we further revealed that the gut microbiota partially mediated the therapeutic benefits of HHXYT and HHXYT contributed to the maintenance of gut microbiota homeostasis. Furthermore, we observed that gut microbiota-metabolized HHXYT contributed to the alleviation of inflammation, oxidative stress, and hormonal imbalance.

Conclusion: This study provided a holistic perspective on the dual mechanism-direct bioactivity and microbiota-dependent effects-underlying the efficacy of HHXYT against PMS, highlighting the importance of host-microbiota interactions in the action of herbal medicines.

Keywords
Blood-absorbed components; Gut microbiota; Hong-Hua-Xiao-Yao Tablet; Metabolomics; Premenstrual syndrome; Transcriptomic.
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