Blood samples were collected 1.5 or 24.5 h after the first treatment and were collected by terminally anaesthetising the mice using isofluraneand performing a cardiac puncture with a 21G needle (Becton Dickinson, Franklin Lakes, NJ, USA) and a heparinised syringe. A volume of approximately 600 µl was collected from each mouse into a 1-ml microcentrifuge tube which had been washed with heparin (Eppendorf, Hamburg, Germany). To separate the serum, the samples were centrifuged at 3,000 rpm at 4°C for 15 min and 300 µl of the supernatant was subsequently transferred to a 1-ml microcentrifuge tube (Eppendorf). Sampled were stored at -20°C until analyzed.
Two liver enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were quantified using a UniCelDxC 600 Synchron Clinical System (Beckman Coulter, Brea, CA, USA) following the manufactures recommend procedures and reagents.
Literature values for the normal range in mice were used to minimize the number of animals used. Specifically, the normal range for AST and ALT in mice are reported to be in the range of 300 ± 100 units/L and 100 ± 50 units/L (Oršolić et al., 2010 (link); Gao et al., 2014 (link)).
Two liver enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were quantified using a UniCelDxC 600 Synchron Clinical System (Beckman Coulter, Brea, CA, USA) following the manufactures recommend procedures and reagents.
Literature values for the normal range in mice were used to minimize the number of animals used. Specifically, the normal range for AST and ALT in mice are reported to be in the range of 300 ± 100 units/L and 100 ± 50 units/L (Oršolić et al., 2010 (link); Gao et al., 2014 (link)).