Forty female Wistar rats (6–8 weeks of age, 160–180 g) were purchased from the Chinese Academy of Military Medical Sciences (Beijing, China). The animals were housed at 25 °C ± 1 °C with 40% ± 5% humidity under 12-h light–dark illumination cycles. The animals were provided food and water ad libitum. The rats were randomly divided into normal, DE, CMC, and CMC + α-MSH groups. The DE, CMC, and CMC + α-MSH groups were injected subcutaneously with 6 mg/mL scopolamine hydrobromide, four times per day for 28 days. This approach has been demonstrated to successfully induce DE in rats11,26. Rats in the CMC group received topical treatment with 0.5% CMC (Allergan, Dublin, Ireland) eye drops, twice per day; rats in the CMC + α-MSH group received 1 × 10−3 mg/mL α-MSH (Millipore US) dissolved in 0.5% CMC eye drops, twice per day. The treatment regimen was initiated on the first day of the modelling protocol. The SIt and corneal fluorescence staining were performed at 7-day intervals. On the 28th day, eyeballs were collected from all rats. These experimental procedures were approved by the Experimental Animal Ethic Committee of Tianjin Medical University (SYXK 2009-0001) and adhered to the tenets of the Association for Research in Vision and Ophthalmology and the Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research.

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