2.1 | Animals
All animal care and study protocols complied with the guidelines of the ethical use of animals and were approved by the Animal Care and Use Committee (Third Military Medical University, Chongqing, China). All efforts were made to minimize animal suffering and reduce the number of animals used. Animal studies are reported in compliance with the ARRIVE guidelines and with the recommendations made by the British Journal of Pharmacology (BJP) (Kilkenny et al., 2010; McGrath & Lilley, 2015). Experimental protocols and design adhere to BJPguidelines (Curtis et al., 2015).
Mice were bred and housed in the laboratory animal room at Chongqing University under specific pathogen-free conditions. Mice were kept in open polypropylene cages with clean chip bedding. The animal room was maintained at a controlled temperature (22 ± 3 °C) and a 12 hr cycle of light and dark. Five mice in each cage were free to drink water and were fed on a regular diet (0.3–0.8% sodium, 1.0–1.8% calcium, 0.6–1.2% phosphorus, Beijing Keao Xieli Feed Corporation, China) unless otherwise indicated. To obtain tissues, all mice were killed by i.p. injection with 2,2,2-tribromoethanol (250 mg·kg−1, Sigma-Aldrich, Cat# T48402).