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TongXie-YaoFang (TXYF) can effectively alleviate the symptoms of diarrhea-predominant irritable bowel syndrome (D-IBS) patients. However, the curative mechanism has not been fully clarified. The study was designed to investigate the effect of TXYF on the colonic ion transport induced by serotonin (5-HT) in D-IBS rats. A method of multiple stress (neonatal maternal separation (NMS) combined with restraint stress (RS)) was used to induce the D-IBS model. The model rats were randomly divided into two groups: NMS + RS group and TXYF-formula group, and the normal control (no handling) rats were classified as NH group. In the NMS + RS group, the change of short-circuit current (ΔIsc) induced by 5-HT was lower than that in the NH and TXYF-formula groups. After removing of the extracellular Cl− or HCO3- or basolateral Na+ or blocking the cystic fibrosis transmembrane conductance regulator (CFTR), Na+-K+-2Cl− cotransporter (NKCC), Na+-HCO3- cotransporter, Cl-/HCO3- exchanger, K+ channel, or Na+/K+-ATPase, respectively, there was no difference in 5-HT-induced ΔIsc among the three groups. These data suggest that TXYF can regulate 5-HT-induced Cl− and HCO3- secretion, possibly mediated by the combined action of CFTR, NKCC, Na+-HCO3- cotransporter, Cl-/HCO3- exchanger, K+ channel, and Na+/K+-ATPase. [ABSTRACT FROM AUTHOR]