Cardioprotective properties of N-terminal galanin fragment (2-15) in experimental ischemia/reperfusion injury

Figures

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  Figure 1. Dose-dependent effect of G on cell viability in response to oxidative stress Treatment of cardiomyoblasts with G prevents H2O2-induced decrease of cell viability in a dose-dependent manner. The H9C2 were pretreated with G (10, 50, 250 nM) for 20 min and then exposed to 400μM H2O2 for 4h. Values are the means ± SEM for three experiments. *P < 0.05 vs H2O2 treatment; **P < 0.01 vs H2O2 treatment; §§§P < 0.001, vs control.

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  Figure 2. Effect of G on hypoxia-induced cell apoptosis (A) Representative fluorescence images of H9C2 cells pretreated with 50 nM G for 20 min and then exposed to normoxia or hypoxia-reoxygenation. Apoptosis was measured by TUNEL assay in H9C2 cells after 16h of hypoxia followed by 3h of reoxygenation. (B) Quantitative analysis of TUNEL-positive cells in H9C2 cells. The arrows indicate apoptotic cells. Values are the means ± SEM from three experiments. ***P < 0.001, vs normoxia (C); §§§P < 0.001, vs hypoxia (H).

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  Figure 3. Effect of G on hypoxia-induced mitochondrial O2- production (A) Representative fluorescence images of H9C2 cells pretreated with G peptide. Mitochondrial O2- formation was assessed by MitoSOX RED in H9C2 cells exposed to 16h hypoxia followed by 3h of reoxygenation. (B) Quantitative analysis of O2- production in H9C2 cells exposed to normoxia or hypoxia-reoxygenation. Values are the means ± SEM from three experiments. *P < 0.05 vs normoxia; ##P < 0.01 vs hypoxia.

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  Figure 4. Effects of peptide G concentrations in KHB on recovery of cardiac output (A) and the index of contractile function intensity (B) at the end of reperfusion. Values are means ± SEM from 8 experiments and are expressed in percentage of the initial value. CO, cardiac output; LVDP×HR, the index of contractile function intensity. The dotted lines show recovery of the indices in control.

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  Figure 5. Effect of G infusion on energy state of isolated rat heart at the end of reperfusion St. st. - steady state; G - 5-min infusion of 225 μ M G after ischemia. (A) Myocardial contents of ATP and ΣAN=ATP+ADP+AMP. (B) Adenylate energy charge (AEC) = (ATP+0.5ADP)/ΣAN. (C) Myocardial contents of PCr and total creatine ΣCr=PCr+Cr. (D) Myocardial content of lactate. Values are the means ± SEM from 8 experiments. *P < 0.05 vs. steady state; #P < 0.05 vs. control.

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  Figure 6. Effects of G administration on necrosis markers in ex vivo and in vivo models of myocardial I/R injury (A) LDH release in myocardial effluent from isolated perfused rat heart. Control - postischemic infusion of KHB; G - postischemic infusion of KHB with 225 μM G. Values are the means ± SEM of 8 experiments and are expressed in IU/g dry wt. for 5-min Langendorff perfusion before or after global ischemia (early reperfusion). * p<0.05 vs. the value before ischemia; # p<0.05 vs. the value in control. Activities of LDH (B) and CK-MB (C) in blood plasma in rats in vivo. St. state - steady state; Control – i.v. administration of saline at the onset of reperfusion; G - i.v. administration of peptide G (1.4 μmol/kg) at the onset of reperfusion. Values are the means ± SEM for 8 experiments.* P<0.05 vs. steady state; # P<0.05 vs. control.

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  Figure 7. Effects of intravenous G administration on myocardial infarct size (MI/AAR, %,) in rats in vivo Black circle corresponds to myocardial infarction in control. Values are means ± SEM from 8 experiments.*P < 0.05 vs. control.