Crf 1 pellet diet

Seven-week-old male and female Sprague-Dawley rats (Crl:CD (SD)) supplied from Charles River Japan Inc. (Tokyo, Japan) were used. Animals were kept in bracket-type stainless steel wire-meshed cages (two or three animals per cage during the study period) at a temperature of 23 ± 3 °C and relative humidity of 55 ± 15% with illumination of 12 h per day from 7 a.m. to 7 p.m. Animals could freely access CRF-1 pellet diet (Oriental Yeast Co., Ltd. (Tokyo, Japan)) and drinking water. Animals were quarantined and acclimated for 2 weeks. Five male and female animals were treated with a single oral dose of 0.5 w/v% methyl cellulose solution (5 mL/kg) using flexible stomach tubes and syringes. Around 0.25 mL/animal of blood was collected via tail vein while conscious and restrained at 0.5, 1, 2, 4, and 8 h after the treatment. In addition, around 2 mL/animal of blood was collected via the abdominal aorta under anesthesia with isoflurane 24 h after the treatment. Blood samples collected in sodium heparin tubes were immediately placed on ice, centrifuged by 10,000 rpm at 4 °C for 2 min to obtain plasma, and stored at − 80 °C until measurement.

Five-week-old male F344 rats were purchased from Charles River (Wilmington, MA, USA). After one week of acclimation, animals were randomly grouped into control or experimental groups. All animal studies were approved by the Animal Care and Use Committee of Kyoto Prefectural University of Medicine (approval number M28-569). The CRF-1 pellet diet was purchased from Oriental Yeast Co., Ltd. (Tokyo, Japan).