Multi-omics Study of the Clinical Efficacy and Mechanism of Linggu Mixture on Functional Constipation Through the Gut Microbiota-Metabolite Axis
Wenwen Zhao, Longhao Tao, Qingfeng Wang, Bianfang YuIntroduction:
Functional Constipation (FC) is a highly prevalent gastrointestinal disorder with inadequate treatment options. Linggu mixture is a Traditional Chinese Medicine (TCM) hospital-prepared formula that has demonstrated clinical efficacy in FC. However, the mechanistic basis of Linggu mixture through the gut microbiota-metabolite axis remains unexplored. The present study systematically evaluated the clinical efficacy of Linggu mixture in FC and elucidated its underlying mechanisms through integrated 16S rRNA sequencing and untargeted metabolomics.
Methods:
The present study involved 22 participants (11 FC patients and 11 healthy controls). The patients received oral Linggu mixture (25 mL, three times daily) for 8 weeks. Clinical outcomes were assessed based on weekly Complete Spontaneous Bowel Movements (CSBMs) and the Wexner Constipation Score. Fecal samples collected pre-treatment (Bm), post-treatment (Am), and from Healthy controls (Hm) underwent 16S rRNA high-throughput sequencing and untargeted ultra- -high performance liquid chromatography–tandem mass spectrometry (UHPLC-MS/MS) metabolomics. Microbiota-metabolite associations were assessed using Pearson correlation analysis.
Results:
Linggu mixture significantly increased weekly CSBMs (1.82 ± 0.75 vs. 4.00 ± 0.89, P < 0.05) and reduced the Wexner Constipation Score by 46.6% (19.90 ± 3.14 vs. 10.63 ± 3.88, P < 0.05). The microbiota analysis revealed significant β-diversity changes (analysis of similarities, R = 0.2438, P = 0.002), with enrichment of beneficial genera such as Akkermansia and Alistipes and a reduction in Ruminococcus. Among 89 differential metabolites identified (Bm vs. Am), indole- 3-propionic acid (IPA) was significantly upregulated, while pro-inflammatory metabolites, including Sphingosine-1-Phosphate (S1P), were downregulated. Correlation analysis revealed that Akkermansia and IPA were significantly positively associated.
Conclusions:
Linggu mixture alleviated FC symptoms and modulated the gut microbiota-metabolite axis in a coordinated manner. The result presented multi-omics-based exploratory evidence supporting a cascade regulatory pathway of microbiota structural remodeling → metabolic pathway regulation → effector molecule action. These results should be interpreted as hypothesis-generating, pending confirmation through larger randomized controlled trials and functional mechanistic studies.