The treatment with sGC stimulator improves survival of hypertensive rats in response to volume-overload induced by aorto-caval fistula

Fig. 1. The experimental design of the whole study depicting the time sequence and experimental manoeuvres; MC – metabolic cage with 12 h urine collection; B – blood collection from the tail vein

Fig. 2. Urinary excretion of cGMP A: throughout the whole observation and B: the difference (Δ) between basal values (day -1) and the end (day 7) in heterozygous Ren-2 transgenic rats (TGR) rats with aorto-caval fistula (ACF) or without (sham) and normotensive sham HanSD rats treated with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined; P > 0.05 (NS); * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001; by 2-way ANOVA with Dunnett's (within each group) and Tukey’s (between groups) multiple comparisons tests; # P ≤ 0.0001 basal values after ACF versus sham operation (day-1) by unpaired T-test

Fig. 3. Angiotensin II (ANG II) A: in plasma and B: in renal tissue in heterozygous Ren-2 transgenic rats (TGR) rats with aorto-caval fistula (ACF) or without (sham) and normotensive sham HanSD rats after 7 day treatment with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined. P > 0.05 (NS); * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001 by one-way ANOVA with Tukey’s multiple comparisons tests; # p < 0.05 versus sham HanSD by one way ANOVA with Tukey’s multiple comparisons tests

Fig. 4. Time course of mean arterial blood pressure (MBP) throughout the whole observation in heterozygous Ren-2 transgenic rats (TGR) rats with aorto-caval fistula (ACF) or without (sham) treated with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined. * P < 0.05 by 2-way ANOVA with Bonferroni's multiple comparisons test; # P < 0.05 versus all other groups 2-way ANOVA with Tukey’s multiple comparisons tests

Fig. 5. Time course of (A) systolic (SBP) and (B) diastolic (DBP) blood pressure throughout 2-week observation in heterozygous Ren-2 transgenic rats (TGR) rats with aorto-caval fistula (ACF) treated with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined. * P < 0.05 versus baseline values on day 0 within each group; # values different between all other groups; $ P < 0.05 values different between BAY41-8543 and both ACEi treated groups of ACF TGR by 2-way ANOVA with Dunnet’s (within each group) and Tukey’s (between groups) multiple comparisons tests. The BP values before the treatment and sham-operated groups are omitted for clarity

Fig. 6. The survival rate in heterozygous Ren-2 transgenic rats (TGR) with aorto-caval fistula (ACF) treated with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined. All groups significantly differ between each other (by log-rank Mantel-Cox test); P > 0.05 (NS); * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001; Control normotensive and sham operated TGR groups were omitted for clarity (the survival in these groups was 90–100% in the end)

Fig. 7. Myocardial protein levels of total Cx43 (A), phosphorylated Cx43 at serine 368 (B), protein kinase C Epsilon (C), and representative western blot images (D) measured in left ventricle collected from heterozygous Ren-2 transgenic rats (TGR) rats with aorto-caval fistula (ACF) or without (sham) and normotensive sham HanSD rats that survived until the end of 7 months treatment with sGC stimulator (BAY41-8543), or with angiotensin-converting enzyme inhibitor (ACEi), alone or combined. Values are normalized to GAPDH and presented as mean ± SEM; Cx43 – connexin 43; PKCε – protein kinase C Epsilon; GAPDH- glyceraldehyde-3-phosphate dehydrogenase; P > 0.05 (NS); * P ≤ 0.05; ** P ≤ 0.01; *** P ≤ 0.001; **** P ≤ 0.0001; by one-way ANOVA and Tukey’s test multiple comparison test; # versus all other groups by one-way ANOVA and Tukey’s test multiple comparison test; (n = 5 in all groups except ACF TGR + BAY41-8543 where only 2 rats survived; note that no rats survived in untreated ACF TGR group)