Sirolimus Enhances Cyclosporine A-Induced Cytotoxicity in Human Renal Glomerular Mesangial Cells
Figure 1
CsA/SRL caused synergistic nephrotoxicity by increasing TGF-β and smad signalling in HMCs. HMCs were grown to confluency and treated with vehicle, 4.2 μM CsA, 1 μM SRL or CsA/SRL, 4.2 μM CsA + 1 μM SRL for 24 hours. (a) Phase contrast micrographs were taken using a CCD camera mounted on a Nikon microscope. Magnification 200X. Cell viability was measured using the resazurin conversion assay. (b) 24-hour TGF-β1 secretion was detected using a TGF-β1 ELISA. (c) Smad activation was measured using a Smad responsive luciferase construct. Smad responsive luciferase values were normalised to a control renilla luciferase construct. Each column represents the mean ± SEM of a minimum of 3 independent experiments performed in duplicate. Data was analysed by ANOVA and comparisons between control and multiple treatment groups were made using the Dunnet posttest. *Indicates statistical difference compared to control. *, **.
