Objective To investigate the effect of regular aerobic exercise on intestinal mucosal tight junction protein claudin-1 in obstructive jaundice mice and the mechanism of guanylate cyclase C (GC-C) pathway in intestinal mucosal barrier function and inflammatory response.   Methods Thirty male Kunming mice were used to establish the obstructive jaundice model and then were randomly divided into 2 groups: regular aerobic exercise group (EX) and model group (M). In addition, the blank group was set up, 15 mice in each group. ELISA, HE staining, intestinal tissue-associated factor immunohistochemical staining, protein detection, Real-time PCR were used in present study in order to explore the effect of the regular aerobic exercise on the obstructive jaundice of the intestinal mucosal.   Results Yellow discoloration of skin in both abdomen and tail was found in mice in M group. The urine color changed into deep yellow and feces appeared light gray.The bile showed cystic dilatation. Serum total bilirubin (TBIL), total bile acid (TBA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and inflammatory factors interleukin (IL)-8, tumor necrosis factor (TNF)-α protein content significantly elevated. Compared with the M group, the protein content of each indicator in the EX group decreased significantly (P<0.01).Microscopically, the hepatic cord arrangement was disordered in group M, and vacuoles, mitosis, and degeneration appeared in large numbers. The intestinal tissue was sparse and uneven, the intestinal gland and muscle layer disappeared or atrophied, and the interstitial layer was edema and inflammatory cells infiltrated. In the EX group, the intestinal mucosa was thickened, and the number of villi significantly increased compared with the M group and the interstitial showed mild edema. Compared with group M, the positive expression, protein content and gene expression of claudin-1, GC-C, guanylin (Gn), uroguanylin (Ugn) and cyclic guanosine monophosphate (cGMP) in EN group increased significantly (P<0.01).   Conclusion Regular aerobic exercise can effectively promote the expression of tight junction protein Claudin-1 in intestinal mucosa, and activate GC-C pathway to reduce the damages of intestinal mucosal barrier function and inflammatory response.