Role of aqueous extract of Sauropus spatulifolius in improving constipation via aquaporin 3/protein kinase B pathway

Objective To explore the efficacy and mechanism of the aqueous extract of Sauropus spatulifolius in treating constipation based on network pharmacology analysis and experiments.

Methods Network pharmacology analysis was conducted using an online platform to investigate the molecular mechanism of Sauropus spatulifolius in treating constipation. Loperamide-induced mouse constipation models and intestinal epithelial cell (IEC) injury models were constructed. Therapeutic effects were evaluated using indicators such as the time to the first black stool, fecal water content, and gastrointestinal transit rate. Annexin V-FITC staining was used to assess apoptosis, JC-1 staining was used to detect mitochondrial membrane potential, chemiluminescence was used to measure adenosine triphosphate (ATP) levels, and western blotting was used to detect the expression of relevant proteins.

Results Network pharmacology analysis revealed that 29 active components in Sauropus spatulifolius targeted 19 genes associated with constipation, with AKT1 identified as one of the key genes. Experimental results demonstrated that the aqueous extract of Sauropus spatulifolius effectively alleviated loperamide-induced constipation symptoms in mice, including weight loss, reduced intestinal motility, prolonged defecation time, and decreased fecal water content. Additionally, the aqueous extract of Sauropus spatulifolius inhibited IEC-6 cell apoptosis, restored mitochondrial membrane potential, and maintained intracellular ATP levels. The therapeutic mechanism involved downregulating the expression of Bcl-2-associated X protein (Bax), cytochrome C, Cleaved-Caspase3, and aquaporin 3 (AQP3), as well as enhancing protein kinase B (Akt) phosphorylation.

Conclusion The aqueous extract of Sauropus spatulifolius effectively ameliorates constipation symptoms in mouse models, and its mechanism may be related to improving intestinal cell energy metabolism, inhibiting IEC apoptosis, and reducing AQP3 expression, suggesting that Sauropus spatulifolius could serve as a potential drug for the clinical treatment of constipation.