Gender Differences in the Progression of Experimental Chronic Kidney Disease Induced by Chronic Nitric Oxide Inhibition

Figures

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  Figure 1. Survival rates of both male and female Control and NAME groups. In this study, 10 animals were included in each experimental group (n = 40). Three male and one female NAME-treated rats died before the end of the study, whereas no mortality was observed in Control groups, regardless of the gender.

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  Figure 2. Representative micrographs of glomerulosclerosis (a) and collapsed glomeruli (b) accessed by PAS staining in groups Control and NAME, male and female. Male NAME rats developed a marked degree of glomerulosclerosis, whereas female NAME rats exhibited a protection of this renal damage feature (a). Glomerular ischemia, represented by the presence of collapsed glomeruli, was also observed in NAME rats, with a higher degree in male than in female animals (b). Accordingly, bar graph representation of percentage of glomerulosclerosis (c) and glomerular ischemia (d) in groups Control and NAME showed that female NAME rats presented renoprotection compared to male NAME animals (Ap < 0.05 versus Control ♂, Bp < 0.05 versus Control ♀, Cp < 0.05 versus NAME ♂). The number of animals studied per group was, respectively, Control ♂: N = 10; Control ♀: N = 10; NAME ♂: N = 7; NAME ♀: N = 9.

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  Figure 3. Representative micrographs of renal cortical interstitial fibrosis (a), evaluated in Masson's trichrome stained slides, and interstitial α-SMA accumulation (b), accessed by immunohistochemistry. Prominent interstitial expansion was observed in both male and female NAME animals, while significant accumulation of interstitial αSMA was seen only in male NAME rats. Accordingly, bar graph representation of percentage of cortical interstitial fibrosis (c) and interstitial α-SMA accumulation (d) showed that female animals submitted to the NAME CKD model were protected from interstitial myofibroblast infiltration when compared to male NAME rats (Ap < 0.05 versus Control ♂, Bp < 0.05 versus Control ♀, Cp < 0.05 versus NAME ♂). The number of animals studied per group was, respectively, Control ♂: N = 5; Control ♀: N = 5; NAME ♂: N = 5; NAME ♀: N = 5.

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  Figure 4. Representative micrographs of immunohistochemical studies in groups Control and NAME: both male and female NAME animals showed a significant increase of renal interstitial macrophage (a) and T-cell (b) infiltration. Additionally, male NAME rats exhibited a statistically increased number of interstitial Ang II+ cells, compared to gender matched Controls (c).

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  Figure 5. Bar graph representation of renal cortical interstitial infiltration by macrophages (a), T-cells (b), and Ang IIα+ cells (c) showed that both male and female NAME rats exhibited a significant increase of renal macrophage and T-cell infiltration. However, NAME females showed statistically less Ang II+ interstitial cells than males subjected to the same treatment (Ap < 0.05 versus Control ♂, Bp < 0.05 versus Control ♀, Cp < 0.05 versus NAME ♂). The number of animals studied per group was, respectively, Control ♂: N = 5; Control ♀: N = 5; NAME ♂: N = 5; NAME ♀: N = 5.

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  Figure 6. Representative micrographs of immunofluorescence double-staining for macrophages (ED1) and angiotensin II (Ang II) in NAME male and female animals. There was no evidence of colocalization of both biomarkers in the same cell (MERGE).

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  Figure 7. Representative micrographs of immunofluorescence double-staining for myofibroblasts (α-SMA) and angiotensin II (Ang II) in NAME male and female animals. There was no evidence of colocalization of both biomarkers in interstitial cells. Colocalization was only constitutively observed in glomerular arterioles and mesangial cells (MERGE).