Figure 1
Light microscopy of immunostained kidney sections A, Neutrophil gelatinase-associated lipocalin (NGAL 200×), endothelin-1 (ET-1, 200×), and catalase (200×) in genetically hypertensive rats in the control group (HT-CTL), hypertensive and hyperglycemic animals (HT-DIAB), hypertensive and hyperglycemic animals treated with macitentan (HT-DIAB+MAC25) for 60 days (final day of the experiment). B, Quantitative analyses of kidney sections stained for NGAL, ET-1, and catalase. Data are reported as %. The significance level for a null hypothesis was set at 5%. **
p < 0.05 compared to the HT-CTL 60 days group and ¥¥
p < 0.05 compared to the HT-DIAB 60 days group (ANOVA followed by the Tukey’s post-hoc test). N= 10 per group.
Figure 2
Renin, angiotensin I (ANG I), and angiotensin II (ANG II) analysis. A, Western blot image of kidney cortex of genetically hypertensive rats (SHR) in the control group (HT-CTL), hypertensive and hyperglycemic animals (HT-DIAB), and hypertensive and hyperglycemic animals treated with macitentan (HT-DIAB+MAC25) for 60 days (final day of the experiment). B,C, and D, Quantitative analyses of immunoblot images were obtained by ImageJ software. Data are reported as means ± SE. The significance level for a null hypothesis was set at 5%. **
p < 0.05compared to the HT-CTL 60 days group and ¥¥
p < 0.05 compared to the HT-DIAB 60 days group (ANOVA followed by the Tukey’s post-hoc test). N = 10 per group.
Figure 3
Analysis of the formation of reactive oxygen species and reactive nitrogen species. A, Quantitative analyses of thiobarbituric reactive substances (TBARS). B, Urinary hydrogen peroxide was measured by a modified ferrous ion oxidation xylenol orange version-2 (FOX-2). C, Nitric oxide (NO) in genetically hypertensive rats in the control group (HT-CTL), hypertensive and hyperglycemic animals (HT-DIAB), and hypertensive animals treated with Macitentan (HT-DIAB+MAC25) for basal, 30, and 60 days. Data are reported as means ± SE. The significance level for a null hypothesis was set at 5%. *
p < 0.05 compared to the HT-CTL 30 days group; ¥P<0.05 compared to the HT-DIAB 30 days group, **
p < 0.05 compared to the HT-CTL 60 days group, and ¥¥ p < 0.05 compared to the HT-DIAB 60 days group (ANOVA followed by the Tukey’s post-hoc test). N = 10 per group.