High-throughput screening identified Epothilone A as a potent GEF-H1 modulator that restores intestinal barrier integrity and improves survival in murine sepsis by suppressing the GEF-H1/librariesRhoA pathway.<h4>Conclusion</h4>This research uncovers the Gzma/GEF-H1/RhoA signalling axis as a pivotal contributor to intestinal barrier dysfunction during sepsis. Here, ARHGEF2 is linked to Sepsis.