Objective To observe the protective effect and mechanism of Daibaijie on intestinal mucosal barrier function. Methods IEC-6 cells were randomly divided into blank control group, LPS group, different dose groups of Daibaijie, LPS +different dose groups of Daibaijie. The cell viability and inflammatory factor levels of IEC-6 cells and IEC-6 cell model induced by LPS were detected after Daibaijie intervention. In addition, Kunming mice were randomly divided into normal control group, LPS group, LPS + Daibaijie high and low dose groups. After one week of continuous administration, the intestinal barrier injury model of mice was established. The content of D-lactate in serum and the mRNA expression levels of TNF-α, IL-1β and IL-6 in colon were detected, pathology of colon were also observed. Results In the range of 30-480 μg/mL, Daibaijie had no significant effect on the cell viability of IEC-6 cells, and could resist the inhibition of LPS on the cell viability of IEC-6 cells, and reduce the levels of IL-1β, TNF-α and IL-6 in IEC-6 cells after LPS induction. Daibaijie could also reduce the levels of D-lactate in LPS induced animal model serum and the mRNA expressions of IL-6, IL-1β and TNF-α in colon, reduce the injury of colon. Conclusion Daibaijie has protective effect on intestinal mucosal barrier function, and the mechanism may be related to improving IEC-6 cell viability and reducing inflammatory response.