Liupao Tea Relieves Constipation by Activating the PI3K/Akt Pathway to Down-Regulate AQP3 and Restore Gut Microbiota

Due to modern lifestyles and dietary changes, the prevalence of constipation is rising and affecting younger populations. Abnormal intestinal water transport is a major factor contributing to constipation, with aquaporins (AQPs) being crucial for water transfer. Previous studies have indicated that dark tea can effectively relieve constipation. Liupao tea, a type of dark tea from Guangxi, is one such tea, but its mechanisms for alleviating constipation remain to be further investigated. This study will use ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with network pharmacology to predict the potential targets and pathways through which Liupao tea alleviates constipation. A constipation rat model will be established by simulating multiple factors that induce constipation in modern life. Hematoxylin and eosin (HE) staining will be used to assess pathological changes in the colon, and immunohistochemistry will measure the expression of inflammatory factors in the colon. High-throughput 16S rRNA sequencing will analyze the effect of Liupao tea on gut microbiota. Real-time quantitative PCR will quantify AQP3 expression in colon tissues, and Western blotting will verify the regulatory effects of Liupao tea on AQP3 protein and the PI3K/Akt pathway. Network pharmacology predictions suggest that Liupao tea may alleviate constipation by modulating the PI3K/Akt pathway. Experimental results show that Liupao tea significantly increases fecal water content in constipated rats and reduces the levels of TNF-α, IL-6, and IL-1β in the colon and stomach tissues. Western blot analysis confirms that Liupao tea significantly enhances Akt phosphorylation and decreases AQP3 expression. Additionally, Liupao tea increases the abundance of gut microbiota in constipated rats. Correlation analysis suggests that Liupao tea may alleviate constipation by improving amino acid metabolism through modulation of the gut microbiota. This study suggests that Liupao tea may alleviate constipation by activating the PI3K/Akt pathway, inhibiting AQP3 expression, and regulating gut microbiota to improve amino acid metabolism.