Objective To observe effect of self-designed drug-loaded nanofilm in preventing postoperative peritoneal cavity adhesion during cholecystectomy in New Zealand white rabbit. Methods The 40 New Zealand white rabbits were randomly divided into blank control group, chitosan group, nanofilm group, and drug-loaded nanofilm group using random number table, the peritoneal cavity adhesions after cholecystectomy at different time (on day 7, 14, 21, and 28) were observed among these 4 groups. Results The adhesion of gallbladder forssa was serious in the blank control group and the adhesion situation had obviously improved among the other three groups, furthermore, the adhesion of the drug-loaded nanofilm group was the slightest. The adhesion score was significantly decreased in the chitosan group, the nanofilm group, or the drug-loaded nanofilm group as compared with the blank control group (P<0.05), which in the drug-loaded nanofilm group was significantly decreased as compared with the chitosan group (P<0.05) or the nanofilm group (P<0.05), which had no significant difference between the chitosan group and the nanofilm group (P>0.05). The nanofilm was degraded on day 14 after surgery and basically completely degraded on day 28 after surgery. The nanofilm degradation points had no significant differences between the nanofilm group and the drug-loaded nanofilm group at different time (P>0.05). Conclusions Drug-loaded nanofilm could prevent postoperative peritoneal cavity adhesion from physical barrier and drug therapy. It provides a new idea for prevention of peritoneal cavity adhesion after general surgery and research and development of new material to prevent peritoneal cavity adhesion in future.