Proximal tubules of the kidneys are one of the most common targets of nephrotoxic drugs and chemicals. Screens to predict nephrotoxic potential of compounds with insights to mechanisms of toxicity facilitate lead optimization, guide structure-activity relationships, minimize risks of clinical nephrotoxicity and therefore are valuable in the process of drug discovery. Innoprot has developed an in vitro High Content Analysis platform to predict the nephrototoxicity. Nephrotoxicity Assay comprises the measurement of the next parameters involved in the renal toxicity:
- Oxidative Stress
- Apoptosis: Caspase 3/7 Activation
- Genotoxicity (DNA damage)
- Cell Viability
- Membrane Damage: LDH Release
We incubate human or animal renal proximal tubule epithelial cells with test-compounds during 24 hours at different concentrations. After treatment, we add probes to measure the oxidative stress, caspase 3-7 activation, and mitochondrial damage. After the probes measurement, we perform the immunoyhistochemistry for H2AX phosphorilation. We finally compare results from the different parameters with negative controls for each parameter and the drug will be refused in the case that the differences for one parameter could be statistically significant in comparison with the negative controls.
Cell line used: Human & Animal Kidney Proximal Tubular Epithelial Cells
Readout: Oxidative Stress, DNA Damage, Caspase Activation, Cell Viability