Integration of network pharmacology and gut microbiota analysis to uncover the molecular mechanisms of Zengye decoction against slow transit constipation.

BACKGROUND

Slow-transit constipation (STC) is the predominant type of functional constipation, and Zengye decoction (ZYD) has been widely used in clinical practice.

PURPOSE: Our aim is to study the improvement effect of ZYD on STC.

STUDY

DESIGN

Network pharmacology and gut microbiota analysis were combined, and the potential underlying molecular mechanism was explored.

METHODS

The chemical components in ZYD were identified by using HPLC-Q-Extractive-MS/MS. The effects of ZYD on STC were subsequently evaluated in diphenoxylate-induced STC rats by measuring body weight changes, discharge time, fecal water content, the number of fecal particles, and the intestinal transport rate. Network pharmacology combined with gut microbiota analysis was subsequently used to explore the possible underlying mechanisms.

RESULTS

A total of 57 compounds, including iridoids, flavonoids, phenylpropanoids, amino acids, organic acids, fatty acids, etc., were identified from ZYD, and iridoids represent the characteristic compounds. ZYD reversed the decrease in body weight in experiment rats, and the feces number and fecal water content of STC rats significantly increased after ZYD treatment (p < 0.05). Moreover, ZYD significantly increased the intestinal transit rate of STC rats (p < 0.05). ZYD protected the integrity of the intestinal mucosal barrier (IMB) via regulation of the ZO-1, Claudin-1, Occludin, MUC-2 and AQP-3 proteins. Network pharmacology analysis revealed that ZYD regulates the MAPK pathway to increase the IMB of STC rats. Furthermore, our results suggested that ZYD regulated the gut microbiota to alter the short-chain fatty acid (SCFA) compositions. SCFAs are closely related to the MAPK signaling pathway.

CONCLUSIONS

Our study suggested that ZYD has potential therapeutic effects on STC by improving the function and integrity of the IMB and that the underlying molecular mechanisms are achieved through the regulation of the intestinal microbiota, short-chain fatty acids (SCFA), and the modulation of the MAPK signaling pathway.