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Placing a physical barrier between the injured site and the adjacent tissues is a very common and highly effective approach to prevent abdominal adhesions in these days. A biodegradable trilayered barrier was fabricated to prevent formation of abdominal adhesions, in which a poly(lactide-co-glycolide)/poly(lactide)-b-poly(ethylene glycol) (PLGA/PLA-b-PEG) electrospun layer was sandwiched between layers of carboxymethyl chitosan (CMCS) sponge. The hydrophilic CMCS sponge layers with glycerin (GL) could adhere to the surface of wound easily, and present great hemostatic capability. The mechanism of the formation of adhesion related to blood clots acting with fibroblast cells was evaluated in detail. The blood clot acted as a "medium" inducing the fibroblast cells growth and proliferation, but had no special attraction on epithelial cells. CMCS sponge layer took away the blood clots during the swelling and dissolution stages. The electrospun layer promoted the growth of epithelial cells, but exhibited inhibition on the adhesion and spread of fibroblast cells, which ensured excellent effect of adhesion prevention. Evaluated by a rat model of sidewall defect-bowel abrasion, significant reductions of postoperative adhesion in its level and occurrence were observed in animals treated by the trilayered barrier.