Male wild type c57bl 6j mice

Male wild-type C57BL/6J mice (6-8weeks old; 20-25 g body weight) were purchased from GemPharmatech Co., Ltd (Nanjing, China), and were housed under controlled environmental condition (12 h light/dark cycles, 24±1°C) with ad libitum access to food and water. Mice were divided into six groups: Sham group, TBI group, TBI+Mino group, TBI+Mino+CYA group, TBI+CYA group and TBI+TGN-020 group. All procedures were performed in accordance with the guidelines from Laboratory Animal Care and Use Committee and Animal Ethics Committee of Wenzhou Medical University. For drug administration, minocycline hydrochloride (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in saline (9 mg/mL) and intraperitoneal (i.p.) injected at the dose of 45 mg/kg 30 min after TBI. CYA (Sangon Biotech, Shanghai, China) and TGN-020 (Sigma-Aldrich, St. Louis, MO, USA) were dissolved in normal saline solution and used for the regulation of AQP4. TGN-020 was i.p. injected at the dose of 200 mg/kg 30 min after TBI, while CYA was i.p. injected at the dose of 100 mg/kg 60 min after TBI (CYA needs delayed injection for its combination use with Mino). These three drugs were injected with same doses at 24 h intervals before mice sacrifice. The efficacy and safety of these doses in mice have been demonstrated previously 27 (link), 28 (link). All efforts were made to minimize the pain and prevent infections.