Rgm 5250

Littermates were randomly assigned to the control and anesthesia groups. The anesthesia group underwent a 4-h exposure to 1.5% sevoflurane in 30% oxygen and 70% room air, and the control group inhaled the carrier gas. Initial experiments did not reveal any sex-based differences in neuroapoptosis, and both female and male littermates were used. A gas analyzer (RGM 5250; Datex-Ohmeda, Louisville, CO, United States) was used to monitor the concentrations of sevoflurane, oxygen, and carbon dioxide. Throughout the experiment, animals were kept in padded acrylic containers equipped with incubators, at a temperature of 37°C.

On P21, rats were randomized to fasting in 30% oxygen (O₂)/70% nitrogen (N₂) or anesthetized with 3% sevoflurane (about 1.5 MAC, Maruishi Pharmaceutical Co., Ltd., Japan, catalog: 95071) plus 30% O₂/70% N₂ for 4 h in control or anesthesia chambers. Inspired anesthetic and oxygen concentrations were monitored using a gas analyzer (RGM 5250; Datex-Ohmeda, Louisville, CO, United States). The temperature in the chambers was maintained via a heating pool at 37 ± 0.5°C. Rats rectal temperature were maintained at 37 ± 0.5°C. Rats for harvest at 2 h, 24 h, and 7 days time points post-treatment were assigned randomly into two study groups: (1) control groups and (2) anesthesia groups. Rats for harvest at the 4 days post-treatment time point were assigned randomly into eight study groups: (1) non-anesthesia + vehicle; (2) non-anesthesia + inhibitor of caspase-3; (3) non-anesthesia + inhibitor of LC3B; (4) non-anesthesia + inhibitor of NF-κB; (5) anesthesia + vehicle; (6) anesthesia + inhibitor of caspase-3; (7) anesthesia + inhibitor of LC3B; (8) anesthesia + inhibitor of NF-κB (n = 6 each group). Rats were sacrificed by intraperitoneal injection of 100 mg/kg pentobarbital (Sinopharm Chemical Reagent Co., Ltd., Shanghai, China, catalog: WS20051129).