To observe the intervention effect of drug-containing serum of Jianpi Yishen Xiezhuo decoction on human colonic epithelial cell inflammation model, and explore the effect of Jianpi Yishen Xiezhuo decoction on intestinal mechanical barrier.Methods 40 SPF SD rats were randomly divided into blank group (n=10), low dose TCM herb enema group (n=10), middle dose traditional Chinese medicine enema group (n=10) and high dose traditional Chinese medicine enema group (n=10). The blank group was not given enema, and the low, middle and high dose groups were given 0.4mL, 0.8mL and 1.6mL traditional Chinese medicine enema at 1∶2∶4 for 30 minutes each time, twice in the morning and evening every day for 5 days. One hour after the last enema, blood was taken from the abdominal aorta, and the upper serum was collected after centrifugation. Human normal colonic epithelial cells (NCM460) were divided into blank group, model group, low-dose TCM herb enema serum group, medium-dose TCM herb enema serum group and high-dose TCM herb enema serum group. Except for the blank group, the other groups were added with 100 μg/mL to induce cell inflammatory injury. The TCM herb enema serum group replaced the cell culture medium with 10% drug-containing serum, and the model group was added with the rat serum of the blank group. After incubation for 24 hours, the supernatant of cells were detected for Claudin-1, Occludin, ZO-1, OD, IL-6, TNF- α. Results Compared with blank serum intervention, TCM herb enema drug-containing serum could enhance the OD value of inflammatory injured cells, but there was no difference among different doses, compared with blank serum intervention, TCM herb enema drug-containing serum could reduce the levels of IL-6 and TNF-α in the supernatant of inflammatory injured cells, and the effect of high dose was more significant. Compared with the blank serum intervention, the drug-containing serum of TCM herb enema can increase the relative expression of Claudin-1, Occludin and ZO-1 of inflammatory injury cells, and the effect of high dose is more significant. Conclusion The drug-containing serum of Jianpi Yishen Xiezhuo decoction can enhance the activity of intestinal epithelial cells and increase the expression of tight junction-related proteins between intestinal epithelial cells, which may help to protect the intestinal mechanical barrier function and reduce local inflammation.