1 M NaOH and 30 μL Doramapimod OPT, and then the reading was performed.

Blank values for GSSG were obtained by reading 100 μL deionized water, 170 μL 0.1 M NaOH plus 30 μL OPT, 15 min after incubation at 25 °C. Fluorometric measures of GSH and GSSG were estimated at 460/40 nm emission wavelengths and 360/40 nm excitation wavelengths. The values of fluorescence were converted to μg/mL by comparison with a correspondent standard curve. Data are shown as mean ± standard error of the mean (SEM) and were analyzed statistically using Instat™ and GraphPad Prism™ software packages. Regression analyses were performed to obtain standard curves of protein, NADH, β-naphthylamine, 4-methoxy-β-naphthylamine, GSH and GSSG. Paired two sided Student’s t-test was performed to compare values of lactate PARP activity dehydrogenase between renal soluble and solubilized membrane-bound fractions. One-way analysis of variance (ANOVA), followed by the Newman–Keuls test when differences were detected, was performed to compare values among groups. Values from a population with equal SDs is a premise of ANOVA, therefore Barlett’s test was applied to verify this hypothesis. In all the calculations, a minimum critical level of p < 0.05 was set. The LD50 corresponded

to 2.08 μg vBj/g body mass and LD50 was used to induce AKI. This value was slightly lower than that found by Ferreira et al. (2005b), that is 2.5 μg vBj/g body mass. Table 1 shows that envenomed mice have reduced hematocrit and plasma urea with increased plasma creatinine and uric acid and unchanged osmolality compared with controls. The increase of creatinine was mitigated by LA, whereas SA restored the normal content of urea in the plasma of animals administered with LD50 of vBj. Both drugs, LA and SA, were similarly efficient to ameliorate the hematocrit and to restore the normal content of uric acid in the plasma of envenomed mice.

Table 2 shows that the LD50 of vBj increased urinary osmolality and creatinine with unchanged uric acid and urea compared with the controls. However, LA associated with LD50 of vBj caused an increase in urinary content of urea compared with the controls. SA decreased the urinary osmolality of Sclareol envenomed mice to lower levels than the controls and was also effective in restoring the normal levels of creatinine in envenomed mice. As shown in Table 3, the LD50 of vBj unchanged the proteinuria, but reduced proteinemia, effect which was not mitigate by both drugs under study. On the contrary, the association of LA with LD50 of vBj caused intense proteinuria. Lactate dehydrogenase activity of the renal cortex and medulla was higher in SF than in MF (Student’s t-test), at levels (data not shown) similar to previously described by Yamasaki et al. (2008). Table 4 shows that the protein content in the SF of the renal cortex was unchanged by the LD50 of vBj.