Figure 1.
Experimental design to measure early perturbations in rat endogenous metabolism, and preliminary studies to determine the dose of gentamicin and time points after exposure to assess injury. Levels of kidney-injury molecule 1 (KIM-1) in urine samples collected at different time windows for rats treated with vehicle (control) or gentamicin (0.50 g/kg) (A) and for rats treated with 0.25 or 0.50 g/kg of gentamicin (B). * Indicates statistically significant differences based on a p value of less than .05. Schematic showing the design of the study, using Sprague Dawley rats exposed to a single dose of 0.50 g/kg of gentamicin under fasting conditions (C). Animals were catheterized 1 week before each experiment and allowed to recover. In Study 1, we administered either gentamicin or vehicle to rats (n = 8 each), and after 13 h, infused them with a combination of 2H/13C tracers to obtain isotope labeling measurements required for metabolic flux analysis. In Studies 2 and 3, we administered gentamicin to rats and observed them for 7 or 13 h (n = 8 in each group with corresponding controls), respectively. We collected samples of liver/kidney tissues and blood/urine after gentamicin exposure (at 7 or 13 h), which we subjected to RNA-sequencing and global metabolic profiling analysis, respectively.