1 Materials and Methods
1.1 Experimental Animals: Female SPF-grade SD rats weighing 180-200g.
1.2 Test Products: Recombinant mussel adhesive protein hydrogel dressing, trademark name Zhan Yan × An Yijia, registration number Xiang Ji Zhu Zhun 20222142096, specifications 0.5 mg/g × 15 g/bottle, batch number 230203, expiration date 20250220, provided by Zhan Yan Biotechnology Co., Ltd.; Collagen gel, specifications 0.5 mg/g × 15 g/bottle, batch number 230202, expiration date 20250202, provided by Zhan Yan Biotechnology Co., Ltd.
1.3 Reagents: Sodium sulfide (specifications 5 g/bottle, Shanghai Aladdin Biochemical Technology Co., Ltd.); Sodium pentobarbital (specifications 5 g/bottle, Sinopharm Chemical Reagent Co., Ltd.). Rat interleukin (IL) 2, IL-6, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α) enzyme-linked immunosorbent assay (ELISA) kits (Jiangsu Enzyme Immune Industry Co., Ltd., catalog numbers MM-0192R1, MM-0190R1, MM-0198R1, MM-0180R1). PI3K, AKT, p-AKT, mTOR antibodies (specifications 10 µL/tube, catalog numbers ab191606, ab179463, ab81283, ab32028, Abcam); p-mTOR antibody (specifications 20 µL/tube, catalog number 5536, CST). Molpure® Cell/Tissue Total RNA Kit (specifications 50 tests, catalog number 19221ES50, Yeasen); PrimeScript RT reagent Kit (specifications 100 Preps, catalog number RR047A, Bioray Biotechnology Co., Ltd.); TB Green™ Premix Ex Taq™ II (Tli RNaseH Plus) (specifications 200 Rxns, catalog number RR820A, Bioray Biotechnology Co., Ltd.).
1.4 Instruments: DH-006 type beauty mole removal pen (Guangzhou Baiyun Han Da Electronics Factory), ultrasonic cell disruptor, model JY92-IIN, Ningbo Xinchi Biotechnology Co., Ltd.; benchtop high-speed refrigerated centrifuge, model H2050R, Xiangyi Group; constant temperature metal bath, model TU-10, Shanghai Yiheng Technology Co., Ltd.; low-temperature high-speed tissue grinder, model KZ-III-F, Servicebio; fluorescence image analysis system, model 5200 Multi, Tanon. Real-time quantitative PCR (RT-PCR) instrument, model QuantStudio™ 3, produced by ThermoFisher Scientific, USA; high-speed low-temperature tissue grinder, model KZ-III-F, produced by Wuhan Seaview Technology Co., Ltd.; thermal cycler, model TCA0096, produced by ThermoFisher Scientific, USA.