Male wistar kyoto rats

Male Wistar Han rats and male Wistar Kyoto rats, at the age of 7 weeks and weighing approximately 180–210 g, were acquired from Charles River (Sulzfeld, Germany). Rats were housed in five animals per standard laboratory cage with free access to food and water and maintained on a 12-h light/dark cycle (lights on at 8 am) under conditions of constant temperature (22 ± 2 °C) and humidity (45 ± 5%). This study was approved by the II Ethical Committee at the Maj Institute of Pharmacology at the Polish Academy of Sciences, Kraków, Poland.

Male Wistar-Kyoto rats (10–11 weeks of age) from Charles River Laboratory (Raleigh, NC) were pair housed in cages containing beta chip bedding in a room under controlled relative humidity (55–65%), temperature (21 °C), and dark/light cycle (12 h each). Rats were provided with water and standard Purina (5001) rat chow (Brentwood, MO) ad libitum unless stated. The animal facility is approved by an Association for Assessment and Accreditation of Laboratory Animal Care. Our animal protocols were approved by the U.S. Environmental Protection Agency’s Institutional Animal Care and Use Committee and all methods were performed in accordance with the relevant guidelines and regulations (NHEERL LAPR #19-05-002). We previously determined that 5–10% of male Wistar Kyoto rats develop spontaneous cardiac hypertrophy^{23 (link)} therefore, heart weights were recorded for all animals at necropsy. To avoid the influence of cardiac hypertrophy-related secondary pulmonary changes, those with a heart/body weight ratio above 20% of normal values were excluded from the study as we have reported in previous study^{8 (link)}. This resulted in n = 6–8 for most endpoints.