Mechanism Study of Yi-Qi-Dao-Zhi Formula in Improving Postoperative Ileus by Targeting the TLR4/NF-κB/MAPK Signaling Pathways

Purpose: Postoperative ileus (POI) commonly impedes recovery after surgical procedures. While the Yi-Qi-Dao-Zhi Formula (YQDZ) has proven its worth in treating premature ovarian insufficiency in clinical settings, the exact therapeutic mechanisms that make it tick continue to fly under the radar. Methods: This investigation employed ultra-performance liquid chromatography combined with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) to pinpoint the key bioactive compounds in YQDZ. For in vivo experiments, a POI mouse model was created through surgical intervention, and gastrointestinal transport function was assessed via charcoal ink experiments. Intestinal histopathology was examined using HE staining. The therapeutic impact of YQDZ on POI was thoroughly assessed using a battery of advanced laboratory techniques, including immunofluorescence staining, immunohistochemical analysis, Western blot procedures, real-time quantitative PCR assays, and ELISA testing. In vitro experiments involved the establishment of an M1-type inflammatory model by inducing mouse mononuclear macrophage RAW264.7 cells with LPS. Additional evaluations encompassed CCK-8 assays, flow cytometry, transcriptomic profiling, real-time PCR, and protein level examinations, all aimed at delving into the mode of action of YQDZ. Results: YQDZ steers M1 macrophage differentiation through the TLR4/NF-κB/MAPK signaling pathway. This action effectively curtails the levels of key inflammatory mediators including TNF-α, IL-1β, and IL-6. Consequently, it acts as a therapeutic agent for inflamed bowels in postoperative ileus mice, helping to restore their gastrointestinal functionality. Conclusion: The therapeutic approach of YQDZ for POI might be connected to the adjustment of the TLR4/NF-κB/MAPK signaling pathway, alongside the inhibition of macrophage polarization.