Sirius Red staining and TUNEL positive apoptotic nuclei in kidney after treatment with ANP.

<p>(A) The red colour of Sirius red staining under the light microscope indicates total collagen deposits, representative images are from renal cortex of each group; (B) Sirius red Score values; (C) Representative micrographs of TUNEL staining in renal cortex and medulla; (D) Apoptotic cells quantification. C-M: Control male; ANP-M: ANP male; C-F: Control female; ANP-F: ANP female. All images are at the same magnification (400X). Scale bar: 30 μm. Value represents mean ± SEM (n = 12) *p <0.01 vs Control male, ^†p<0.01 vs Control female.</p

Effect of chronic treatment with ANP on the enzymes involved in renal oxidative state of male and female SHR.

<p>CAT: Catalase; SOD: superoxide dismutase; GPx: glutathione peroxidase.</p><p>Value represents mean ± SEM (n = 12).</p><p>*p <0.01 vs control male</p><p>†p<0.01 vs control female</p><p>Effect of chronic treatment with ANP on the enzymes involved in renal oxidative state of male and female SHR.</p

Effects of ANP treatment on renal NOS.

<p>(A) Nitric oxide synthase (NOS) activity; (B) Representative blots of eNOS and β-actin; (C) Quantification of eNOS bands. All experiments were performed in triplicate. β-actin was used as a loading control and data are normalized to actin expression. White spaces demarcate noncontiguous gel lanes. Data are mean ± SEM (n = 12). *p<0.01 vs Control male, ^†p<0.01 vs Control female.</p

Sirius Red staining in left ventricle after treatment with ANP.

<p>The red colour of Sirius red staining under the common microscope indicates total collagen deposits, representative images are from hearts of each group. (<b>A</b>): Control male; (<b>B</b>): ANP male; (<b>C</b>): Control female; (<b>D</b>): ANP female. All images are in the same magnification 400X. Scale bar  = 30 µm.</p

ANP treatment effects on SBP, NOx, cardiac NOS activity and eNOS expression.

<p>(<b>A</b>) SBP: systolic blood pressure; (<b>B</b>) NOx: Nitrites and nitrates excretion; (<b>C</b>) NOS (nitric oxide synthase) activity; (<b>D</b>) Representative blot of eNOS and β-actin and quantification of the bands of eNOS. Data are mean ± SEM (n = 10). *p<0.01 vs. Control male, # p<0.01 vs Control female.</p

Effects of chronic treatment with ANP on oxidative stress in left ventricle of male and female SHR.

<p>GSH: glutathione; TBARS: thiobarbituric acid reactive substances; CAT: Catalase; SOD: superoxide dismutase; GPx: gluthatione peroxidase.</p><p>Data are mean ±SEM (n = 10).</p><p>*p<0.01 vs. Control male;</p>#<p>p<0.01 vs. Control female.</p

Effects of chronic treatment with ANP on cardiac fibrosis and apoptosis in male and female SHR.

<p>SR: Sirius red; TGF-β: transforming growth factor beta.</p><p>Data are mean ±SEM (n = 10).</p><p>*p<0.01 vs. Control male;</p>#<p>p<0.01 vs. Control female.</p

Immunohistochemistry staining for TGF-β in left ventricle after treatment with ANP.

<p>Representative micrographs of immunostained TGF-β in the hearts from each group. (<b>A</b>): Control male; (<b>B</b>): ANP male; (<b>C</b>): Control female; (<b>D</b>): ANP female. All images are in the same magnification 400X. Scale bar  = 30 µm.</p

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AQP2 mRNA levels in the renal outer medulla.

<p>AQP2 mRNA levels are expressed as relative values from control untreated rats. The following formula was applied: ΔΔCT = (<i>CTAQP</i>2−<i>CTGAPDH</i>) <i>experimental</i>−(<i>CT AQP</i>2−<i>CT GAPDH</i>)<i>control untreated rats</i>. Two-way ANOVA showed a statistically significant (p<0.001) interaction between the effects of Diabetes and L-Arg treatment on AQP2 mRNA expression. Bonferroni’s post- tests: *p<0.05 vs. control untreated rats; ***p<0.001 vs. control untreated rats; &&&p<0.001 vs. diabetic untreated rats. Data are mean ± SEM (n = 6).</p