CI-1040 inhibits phospho-ERK1/2 expression after SNx and upregulates p38, cJun and PAI-1.

<p>(<b>a</b>) terminal kidney homogenates were western blotted for pERK1/2, phospho-p38 (p-p38), phospho-cJun (p-c-Jun) and PAI-1 with calnexin and total ERK1/2 acting as loading controls. (<b>b</b>) Densitometry values plotted with data representing mean +/- SEM, n = 6–11 per group. Statistical significance determined by one-way ANOVA. ***p<0.0001.</p

CI-1040 has no effect on blood pressure, kidney function and albuminuria after SNx.

<p>CI-1040 does not improve kidney function in the SNx rat; systolic blood pressure (SBP) (<b>a</b>), urinary albumin excretion (<b>b</b>), serum creatinine (<b>c</b>) and creatinine clearance (<b>d</b>) after 18 weeks treatment with either CI-1040 60mg/kg/d (closed triangles) or vehicle (closed circles) (n = 11 per group). Vertical bars indicate ± SEM, * p<0.05, ** p<0.001, *** p<0.0001 vs sham controls (open circles).</p

Myofibroblast number after treatment with CI-1040.

<p>Myofibroblast number as indicated by α-SMA staining. Representative sections of glomeruli (400x) from terminal kidney tissue (90 day) stained with α-SMA obtained from (<b>a</b>) sham (<b>c</b>) SNx and (<b>e</b>) SNx plus CI-1040 60mg/kg/day rats. Group data are quantified in (<b>g</b>). Representative sections of the tubulointerstitium (200x) from terminal kidney tissue (90 day) stained with α-SMA obtained from (<b>b</b>) sham (<b>d</b>) SNx and (<b>f</b>) SNx plus CI-1040 60mg/kg/day rats. Group data are quantified in (<b>h</b>). Bars represent mean ± SEM, n = 7–11 per group. * p<0.05, *** p<0.001 vs sham controls.</p

CI-1040 inhibits pERK1/2 activation and proliferation in rat fibroblasts.

<p>NRK49F cells were serum starved overnight together with increasing concentrations of the MEK inhibitor CI-1040 prior to stimulation with 10% foetal bovine serum. pERK1/2 expression was assessed by western blotting with calnexin as a loading control (1a). CI-1040 treatment leads to a dose-dependent reduction in pERK1/2 expression as a percentage of control (n = 3). Cell proliferation as assessed by BrdU ELISA (1b) shows CI-1040 at doses between 100nM and 10,000nM has no significant effect on cell proliferation (closed circles). Viability assays (1b, closed triangles) determined CI-1040 was cytotoxic at doses higher than 10,00nM (assay performed 3 times in triplicate. V to refers vehicle only. + refers to FBS-stimulated cells and–refers to non-stimulated cells.</p