Objective To investigate the effects of rhubarb combined with glutamine on repair of gut mucosal barrier injury after gastrointestinal surgery in rats. Methods Fifty male healthy Sprague-Dawley(SD)rats were randomly divided into five groups:control(C group),enteral nutrition(EN group),rhubarb with EN(R-EN group), glutamine with EN(G-EN group)and rhubarb combined with glutamine and EN(R-G-EN group)groups(each n=10). The rats of C group received no treatment and were fed normally. Cecectomy and gastrostomy were performed in EN,R-EN,G-EN and R-G-EN groups,and in 24 hours after surgery,they were interfered with the use of amoxicillin 50 mg/d and metronidazole 20 mg twice a day. From the first day after operation,each group was treated with the same prescription of enteral isocaloric and isonitrogenous nutrition for consecutive 7 days. Seven days later, under anaesthesia,laparotomy was performed,and a specimen of 1 cm intestinal segment was taken from the terminal 5 cm long ileum for all the groups;under the light microscope,the morphological examination of intestinal mucosal epithelial tight junctions(including villus height and mucosal thickness)was carried out. For each group,enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of plasma citruline and D-lactate,and the serum concentration of TNF-αwas detected by utilizing double antibody ABC-ELISA. Results The levels of D-lactate and TNF-αin EN,R-EN,G-EN and R-G-EN groups were obviously higher than those in C group(all P<0.01). The level of plasma citruline(μmol/L:16.13±0.58 vs. 14.52±0.28)of R-G-EN group was remarkably higher than that in R-EN group(P<0.05),and its levels of serum D-lactate(mmol/L:3.12±0.56 vs. 3.62±0.52)and TNF-α(ng/L:2.82±0.62 vs. 3.54±0.49)were markedly lower than those of G-EN group(both P<0.05). The mucosal villus height(μm:126.50±6.31 vs. 175.67±11.91),mucosal thickness(μm:223.67±11.94 vs. 277.00±14.06)and the level of plasma citruline(μmol/L:13.25±0.36 vs. 15.67±0.55)in EN group were obviously lower than those of C group(all P<0.01);the above indexes of R-EN,G-EN and R-G-EN groups were higher than those of EN group (P<0.05 or P<0.01),but in the comparisons of the villus height and mucosal thickness among the three groups there were no statistical significant differences(all P>0.05). Conclusion The application of rhubarb combined with glutamine can protect gut mucosa,reduce the level of inflammatory reaction and elevate the level of plasma citruline, that is beneficial to the repair of gut mucosal barrier injury and its functional recovery after gastrointestinal surgery in rats.