Intrauterine adhesion (IUA) is a disease characterized by endometrial fibrosis caused by injury to the endometrium. In the present study, decellularized and lyophilized human amniotic membrane (DL-AM) material was transplanted in a rat model to explore the preventive effect against IUA. A total of 24 Sprague Dawley rats were randomly divided into an IUA (n=12) group and an IUA + DL-AM (n=12) group. To establish the model, the endometrium of the left uterus was scraped, while that of the right uterus was used as a control. In the IUA group, scraped uteri were sutured without any other treatment, whereas DL-AM was transplanted onto the scraped uteri in the IUA + DL-AM group. Uteri were resected for histological and immunohistochemical evaluation at 3, 7, 14 and 28 days after surgery. The results confirmed the development of IUA, which was accompanied by an increase in the rate of fibrotic area. Integral optical density (IOD) values of connective tissue growth factor (CTGF) were elevated in the IUA group, while matrix metalloproteinase-2 (MMP-2) decreased relative to the control group (P<0.05). After DL-AM transplantation, the IOD value of CTGF dropped, while MMP-2 increased compared with the IUA group (P<0.05). However, compared with that in the control group, the IOD value of CTGF was still higher, whereas MMP-2 was still lower in the IUA + DL-AM group (P<0.05). Furthermore, no evidence of endometrial regeneration was detected in both the IUA and IUA + DL-AM groups. Overall, these results indicated that in the rat model of IUA, transplantation of DL-AM had the potential to prevent the formation of fibrosis to a certain extent and may thus be an alternative strategy for managing the condition.