Inhibition of DDX5-Mediated G-Quadruplex Unwinding in the β-Catenin 5′-UTR by Magnesium Isoglycyrrhizinate Alleviates Chemotherapy-Induced Intestinal Injury

Chemotherapy-induced gastrointestinal toxicity, particularly intestinal barrier disruption and diarrhea, is a major dose-limiting adverse effect with unclear mechanisms. Here, we identify magnesium isoglycyrrhizinate (MIG) as a novel therapeutic agent that ameliorates 5-fluorouracil (5-FU)- and irinotecan-induced intestinal injury by enhancing epithelial barrier integrity. Using MIG as a molecular probe, we revealed a pathogenic mechanism underlying chemotherapy-induced barrier damage. Integrated chemoproteomics (limited proteolysis-mass spectrometry and thermal proteome profiling) analyses revealed direct binding of MIG to the RNA helicase DDX5 (DEAD-box helicase 5), whose expression is markedly increased upon 5-FU-induced injury. Mechanistically, DDX5 destabilizes the G-quadruplex (G4) structure in the <i>CTNNB1</i> (β-catenin) 5' untranslated region, suppressing β-catenin production and compromising barrier function. Crucially, MIG acts as a novel DDX5 inhibitor that blocks G4 unwinding, thereby restoring β-catenin expression and barrier integrity in a dose-dependent manner. In vivo, MIG outperforms the canonical DDX5 inhibitor supinoxin in mitigating intestinal damage. Taken together, the results of our study not only establish MIG as a promising therapeutic candidate but also delineate the DDX5/<i>CTNNB1</i> axis as a targetable pathway for treating chemotherapy-induced barrier dysfunction.