This study aimed to explore the influence of the combination of rhubarb polysaccharide and semen crotonis pulveratum on lymphocyte homing in the intestinal mucosa of ulcerative colitis (UC) rats. A rat colitis model was induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Rats were allocated to the control group, the model group, the salazosulfapyridine group, the semen crotonis pulveratum+ rhubarb polysaccharide high dose group (B+R80 group), the moderate group (B+R60 group), and the low group (B+R40 group) (all n=10). Flow cytometry detected CD3+, CD4+, CD8+ and the CD4+/CD8+ ratio in peripheral blood and the lamina propria of the intestinal mucosa. Mucosal addressin cell adhesion molecule-1 (MAdCAM-1) mRNA in intestinal mucosal tissues was monitored with RT-PCR. Lymphocyte function-related molecule (alpha L beta 2) and intercellular adhesion molecule-1 (ICAM-1) were detected by immunohistochemistry in colon tissues. Compared with the model group, CD8+ levels in the peripheral blood from the B+R40, B+R60, and B+R80 groups increased (P<0.05), and the CD4+/CD8+ ratio decreased (P<0.01). CD4+ and CD4+/CD8+ ratio in the intestinal mucosa were increased (P<0.01), and the difference was most notable in the B+R40 group. The colon macroscopic damage index score decreased to different degrees (P<0.05), histopathologic score decreased (P<0.01), and the expression of MAdCAM-1 decreased. The expression of ICAM-1 was significantly decreased (P<0.05), but similar between groups (P>0.05). alpha L beta 2 expression was significantly decreased in the B+R80 and B+R40 groups (P<0.01). Rhubarb polysaccharide and semen crotonis pulveratum in combination decreased damage to the intestinal mucosa by regulating homing of lymphocytes in UC rats to improve the immune function of the mucosa.