Animal model of CS

Male Wistar rats weighing 250–300 g were obtained from Japan SLC (Shizuoka, Japan) and housed in a room maintained at a temperature of 23 ± 3 °C and a relative humidity of 55 ± 15% under a 12:12-h light/dark cycle, with free access to food and water. Animal experiments were carried out according to the guidelines for animal use and approved by the Life Science Research Center of Josai University (approval nos. {"type":"entrez-nucleotide","attrs":{"text":"H26027","term_id":"895150","term_text":"H26027"}}H26027, {"type":"entrez-nucleotide","attrs":{"text":"H26028","term_id":"895151","term_text":"H26028"}}H26028, {"type":"entrez-nucleotide","attrs":{"text":"H27025","term_id":"897015","term_text":"H27025"}}H27025, and {"type":"entrez-nucleotide","attrs":{"text":"H27027","term_id":"897017","term_text":"H27027"}}H27027). Anesthesia was induced by intraperitoneal injection of sodium pentobarbital (50 mg/kg body weight). Body temperature was maintained throughout the experiment using a heating pad. The CS model was established as previously reported [12]. Briefly, a rubber tourniquet was applied to the bilateral hind limb of each rat, wrapped five times around a 2.0 kg metal cylinder, and the end of the band was glued. After a compression of 5 h, the compression was released by cutting the band and removing the tourniquet.