TRIP6 is a zyxin family member that serves as an adaptor protein to regulate diverse biological processes. In prior reports, TRIP6 was shown to play a role in regulating inflammation by inducing the activation of the nuclear factor-κB (NF-κB) signaling cascade, though its specific role in vivo in inflammatory contexts remains to be clarified. Herein, we therefore employed TRIP6-deficient (TRIP6 −/− ) mice in order to explore the mechanistic importance of TRIP6 in a dextran sodium sulfate (DSS)-induced model of murine colitis. Findings Wild-type (TRIP6 +/+ ) mice developed more severe colitis following DSS-mediated disease induction relative to TRIP6 −/− mice, as evidenced by more severe colonic inflammation and associated crypt damage. From a mechanistic perspective, TRIP6 expression in wild-type mice was significantly elevated in the period of DSS treatment; TRIP6 deficiency was associated with the inhibition of IκBα degradation, thus suppressing NF-κB signaling activity and impairing the secretion of proinflammatory cytokines such as TNFα and IL-6. Conclusions Together, these in vivo data demonstrate that TRIP6 serves as a positive regulator of DSS-induced colitis by regulating inflammatory NF-κB signaling, highlighting its therapeutic promise as a protein that theoretically can be targeted to prevent or treat colitis.