A Slow Hydrogen Sulfide Donor GYY-4137 Partially Improves Vascular Function in Spontaneously Hypertensive Rats Fed a High-Fat Diet

Figures

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  Figure 1. The systolic blood pressure of rats. The following groups were used: the control SHR (SHR, n = 8), SHR receiving HFD (SHR + HFD, n = 7), and SHR receiving HFD and 266 µmol/kg/day GYY-4137 (SHR + HFD + GYY, n = 8) groups. The data are shown as the means ± SEMs. The statistical analysis was performed by two-way ANOVA with the Bonferroni post hoc correction. N—number of rats. *** p < 0.001 compared with the SHR group (17th week), ++ p < 0.01 compared with the SHR + HFD group (17th week), <<< p < 0.001 compared with the SHR group (14th week).

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  Figure 2. Contractile responses induced by noradrenaline in the thoracic aorta (A) and mesenteric artery (B) and endothelium-dependent vasorelaxant responses induced by acetylcholine in the thoracic aorta (C) and mesenteric artery (D). Arteries were isolated from control rats (SHRs), SHRs fed the HFD (SHR + HFD), and SHRs fed the HFD and treated with GYY-4137 (SHR + HFD + GYY). Number of rats: thoracic aorta—SHR group n = 6, SHR + HFD group n = 6–7, and SHR + HFD + GYY group n = 7–8; and mesenteric artery—SHR group n = 7, SHR + HFD group n = 6, and SHR + HFD + GYY group n = 6–7. The results are presented as the means ± S.E.Ms. Statistical analyses were performed by two-way ANOVA with the Bonferroni post hoc correction. * p < 0.05, and *** p < 0.001 compared with the SHR group, and + p < 0.05 compared with the SHR + HFD group.

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  Figure 3. The effects of NG-nitro-L-arginine methyl ester (LN) on the contractile responses of the thoracic aorta induced by noradrenaline in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of NO participation was determined via an AUC evaluation (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 6; SHR + HFD n = 5; and SHR + HFD + GYY n = 7. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. LN within the relevant group; +++ p < 0.05 vs. SHR LN+.

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  Figure 4. Effects of NG-nitro-L-arginine methyl ester (LN) on the relaxation responses of the thoracic aorta induced by acetylcholine in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of NO participation was determined via an AUC evaluation (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 6; SHR + HFD n = 6; and SHR + HFD + GYY n = 6. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05, *** p < 0.001 vs. LN within the relevant group; + p < 0.05; +++ p < 0.001 vs. SHR LN+.

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  Figure 5. Effects of NG-nitro-L-arginine methyl ester (LN) on the contractile responses of the mesenteric artery induced by noradrenaline in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of NO participation was determined via an AUC evaluation (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 6; SHR + HFD n = 6; and SHR + HFD + GYY n = 7. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or a paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05 vs. LN within the relevant group; + p < 0.05 vs. SHR LN+.

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  Figure 6. Effects of NG-nitro-L-arginine methyl ester (LN) on relaxation responses of the mesenteric artery induced by acetylcholine in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of NO participation was determined via an AUC evaluation (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 7; SHR + HFD n = 6; and SHR + HFD + GYY n = 6. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or a paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the relevant group without LN treatment.

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  Figure 7. Effect of DL-propargylglycine (PPG) on the contractile response of the thoracic aorta induced by noradrenaline in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of H2S participation was evaluated via the AUC (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 7; SHR + HFD n = 6; and SHR + HFD + GYY n = 7. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05 and *** p < 0.001 compared within the relevant group without PPG treatment.

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  Figure 8. Effects of DL-propargylglycine (PPG) on the relaxation responses of the thoracic aorta induced by acetylcholine in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of H2S participation was evaluated via the AUC (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 6; SHR + HFD n = 6; and SHR + HFD + GYY n = 6. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05 and *** p < 0.001 compared within the relevant group without PPG treatment; + p < 0.05 compared with the SHR + PPG group.

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  Figure 9. Effect of DL-propargylglycine (PPG) on the contractile response of the mesenteric artery induced by noradrenaline in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of H2S participation was evaluated via the AUC (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 7; SHR + HFD n = 6; and SHR + HFD + GYY n = 7. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with the Bonferroni post hoc correction. * p < 0.05 and ** p < 0.01 compared with the relevant group without PPG treatment.

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  Figure 10. Effects of DL-propargylglycine (PPG) on the relaxation responses of the thoracic aorta induced by acetylcholine in control SHRs (A), SHRs fed the HFD (B), and SHRs fed the HFD and treated with GYY-4137 (C). The level of H2S participation was evaluated via the AUC (D). White column—before the addition of the inhibitor; hatched column—after the addition of the inhibitor. The number of arterial segments per group was as follows: SHR n = 6; SHR + HFD n = 6; and SHR + HFD + GYY n = 6. The results are presented as the means ± S.E.Ms. The statistical analysis was performed by two-way ANOVA (line graphs) or paired Student’s t test/one-way ANOVA (AUC) with Bonferroni post hoc test.

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  Figure 11. Total NOS activity in aortic tissue. Arteries were isolated from control rats (SHRs), SHRs fed the HFD (SHR + HFD), and SHRs fed the HFD and treated with GYY-4137 (SHR + HFD + GYY). The numbers of samples from the rats in each group were as follows: SHR n = 8, SHR + HFD n = 6, and SHR + HFD + GYY n = 6. The data are presented as the means ± S.E.Ms. The statistical analysis was performed by one-way ANOVA. * p < 0.05 compared with SHRs.

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  Figure 12. Protein expression of endothelial NOS (A), inducible NOS (B), the H2S-producing enzyme CSE (C), CBS (D), and TNFα (E) in aortic tissue. Arteries were isolated from control rats (SHRs), SHRs fed the HFD (SHR + HFD), and SHRs fed the HFD and treated with GYY-4137 (SHR + HFD + GYY). Endothelial NOS—eNOS; inducible NOS—iNOS; CSE—cystathionine γ-lyase; CBS—cystathionine β-synthase; TNFα—tumor necrosis factor alpha. The numbers of samples from rats in each group (for each protein) were as follows: SHR n = 6, SHR + HFD n = 6, and SHR + HFD + GYY n = 6. The data are presented as the means ± S.E.Ms. The statistical analysis was performed by one-way ANOVA. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the SHR group; + p < 0.05, ++ p < 0.01, and +++ p < 0.001 compared with the SHR + HFD group.