Effect of (-)-epicatechin on the modulation of progression markers of chronic renal damage in a 5/6 nephrectomy experimental model

Figures

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  Fig. 1. Systolic blood pressure (SBP) at 14 days post-surgery. There is an increase in SBP in the group with nephrectomy 5/6 (Nf) vs Sham. A dose dependent decrease in SBP was found in the Nf treated with Epi (Nf + Epi) vs (Nf) without treatment. Data are presented as mean ± sem (n = 11). Analysis performed with one-way ANOVA with Post-Hoc test Tukey. ***p < 0.001, NS: not significant.

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  Fig. 2. Determination of the concentration of serum creatinine and urea at 14 days post-surgery. A) There is an increase in creatinine in the group with nephrectomy 5/6 (Nf) vs Sham. A dose dependent decrease in creatinine was found in the Nf treated with Epi (Nf + Epi) [0.01 to 1 mg/kg of body weight/day] vs (Nf) without treatment. B) There is an increase in Urea in the group with nephrectomy 5/6 (Nf) vs Sham. A dose dependent decrease in urea was found in the Nf treated with Epi (Nf + Epi) [0.01 to 1 mg/kg of body weight/day] vs (Nf) without treatment. Data are presented as mean ± sem (n = 11). Analysis performed with one-way ANOVA with Post-Hoc test: Tukey. **p < 0.01, ***p < 0.001, NS: not significant.

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  Fig. 3. Serum concentration of tetrahydrobiopterin (BH4) at 14 days post-surgery. There is a decrease of BH4 concentration in the nephrectomy 5/6 (Nf) without treatment group vs Sham. In contrast there is an increase in BH4 concentration in the Nf treated with Epi (Nf + Epi) groups [0.01 to 1 mg/kg of body weight/day]. Data are presented as mean ± sem (n = 11). Analysis performed with one-way ANOVA with Post-Hoc test: Tukey. ***p < 0.001, NS: not significant.

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  Fig. 4. Representative images of Hematoxylin & Eosin staining of histological sections of kidney (8μm), obtained from sham surgery groups treated with vehicle (Sham + V) (Fig. 4A [40x] and B [100x]), with nephrectomy 5/6 (Nf) without treatment (Fig. 4C [40x] and D [100x]) and with Nf treated with Epi (Nf + Epi) at the doses of 0.01 mg/kg of body weight/day (Fig. 4E [40x] and F [100x]), 0.1 mg/kg of body weight/day, (Fig. 4G [40x] and H [100x]) and 1 mg/kg of body weight/day (Fig. 4I [40x] and J [100x]).

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  Fig. 5. Analysis of the expression of angiotensin II type 1 receptor (AT1-R) in renal tissue analyzed by western blot, at 14 days post-surgery. The AT1-R/tubulin index is reported in arbitrary units (A.U.). Significant increase in the expression of ATR-1 in the group with the nephrectomy 5/6 (Nf) vs Sham. There is no change in the group treated with Epi 1 mg/kg of body weight/day as compared with Sham group. Data are presented as mean ± sem. Analysis performed with one-way ANOVA with Post-Hoc test: Tukey. *p < 0.05, NS: not significant.

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  Fig. 6. Analysis of NOX-4 expression in renal tissue analyzed by western blot, at 14 days post-surgery. The index NOX-4/tubulin is reported in arbitrary units (A.U.). The expression of NOX-4 was significantly increased in the group with the nephrectomy 5/6 (Nf). In contrast, in the Nf group treated with Epi (Nf + Epi) at 1mg/kg of body weight/day NOX-4 the expression remained without significant difference vs Sham. Data are presented as mean ± sem. Analysis performed with one-way ANOVA with Post-hoc test: Tukey. *p < 0.05, NS: not significant.

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  Fig. 7. Dot blot analysis of the formation of 3-nitrotyrosine in the renal tissue at 14 days post-surgery. 3-nitrotyrosine/tubulin index was reported in arbitrary units (A.U.). A significant increase in the formation of 3-nitrotyrosine in the group with Nf vs Sham group was noted. The group with Nf treated with Epi at 1 mg/kg of body weight/day showed a decrease in 3-nitrotyrosine without significant differences vs Sham group. Data are presented as mean ± sem. Analysis performed with one-way ANOVA with Post-Hoc test: Tukey. *p < 0.05, NS: not significant.