Figure 1 Empa and Cariporide effects during IR in insulin-perfused hearts. Hearts were subjected to IR and 1 µM Empa, ... Figure 1 Empa and Cariporide.

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Presentation transcript:

Figure 1 Empa and Cariporide effects during IR in insulin-perfused hearts. Hearts were subjected to IR and 1 µM Empa, ... Figure 1 Empa and Cariporide effects during IR in insulin-perfused hearts. Hearts were subjected to IR and 1 µM Empa, 10 µM Cariporide, or vehicle in the presence of insulin (A). All three conditions display similar time points of ischaemic contracture development (B + C). No changes in any IR injury parameters were detected by Empa or Cariporide, including global infarct size (D), RPP recovery at t = 2 h reperfusion relative to baseline RPP (E), end-diastolic pressure (F), and total LDH release during reperfusion (G). Vehicle n = 5, Empa n = 6, and Cariporide n = 6. Scale bar = 0.5 mm. LDH AUC, Lactate dehydrogenase activity area under the curve during reperfusion, normalized to heart wet weight; Pdia, diastolic pressure; RPP, rate pressure product; TOC, time of onset contracture. Statistical testing by one-way ANOVA. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 2 Empa and Cariporide effects during IR in non-insulin-perfused hearts. Hearts were subjected to IR and 1 µM ... Figure 2 Empa and Cariporide effects during IR in non-insulin-perfused hearts. Hearts were subjected to IR and 1 µM Empa, 10 µM Cariporide, or vehicle in the absence of insulin (A). Both Empa and Cariporide delayed ischaemic contracture development (B + C), whereas only Cariporide administration proceeded to improvements in IR injury, measured as global infarct size (D), RPP recovery at t = 2 h reperfusion relative to baseline RPP (E), end-diastolic pressure (F), and total LDH release during reperfusion (G). Vehicle n = 14, Empa n = 13, and Cariporide n = 15. Scale bar = 0.5 mm. LDH AUC, Lactate dehydrogenase activity area under the curve during reperfusion, normalized to heart wet weight; Pdia, diastolic pressure; RPP, rate pressure product; TOC, time of onset contracture. *P < 0.05, **P < 0.01 by one-way ANOVA with Dunnett’s post hoc vs. vehicle except for Figure 2F (Pdia), which was tested by the Kruskal–Wallis test and the Mann–Whitney U test with Bonferroni correction. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 3 Empa effects under NHE1 inhibiting conditions during IR in non-insulin-perfused hearts. Hearts were subjected ... Figure 3 Empa effects under NHE1 inhibiting conditions during IR in non-insulin-perfused hearts. Hearts were subjected to IR and 2.5 µM Eniporide, with or without 1 µM Empa in the absence of insulin (A). Empa did not further delay ischaemic contracture onset (B + C). No differences between groups were observed in global infarct size (D), RPP recovery at t = 2 h reperfusion relative to baseline RPP (E), end-diastolic pressure (f), and total LDH release during reperfusion (G). N = 6/condition. Scale bar = 0.5 mm. LDH AUC, Lactate dehydrogenase activity area under the curve during reperfusion, normalized to heart wet weight; Pdia, diastolic pressure; RPP, rate pressure product; TOC, time of onset contracture. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 4 Presence of insulin results in increased baseline glycogen, ischaemic glycogen depletion, G6P, lactate ... Figure 4 Presence of insulin results in increased baseline glycogen, ischaemic glycogen depletion, G6P, lactate accumulation, and mtHKII dissociation at end-ischaemia. Hearts were subjected to 20 min baseline (AΙ) and 25 min ischaemia (AΙΙ). Insulin extensively delayed contracture development during ischaemia (B). Cardiac glycogen content (C), G6P (D), and lactate (E) were determined at baseline (n = 3/4 per group) and end-ischaemia (n = 6 per group) for insulin- and non-insulin-perfused hearts. Typical examples of HKII and VDAC western bands for mitochondrial compartment (F). MtHKII protein content normalized to VDAC (G + H) and HK activity normalized to CS activity (I + J) in baseline and end-ischaemic hearts, respectively. G6P, glucose-6-phosphate; HKII, hexokinase 2; VDAC, voltage-dependent anion channel. *P < 0.05, **P < 0.01, and ***P < 0.001 by one-way ANOVA with Bonferroni post hoc. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 5 Insulin does not affect NHE activity or the ability of Empa and Cariporide to inhibit NHE in isolated mouse ... Figure 5 Insulin does not affect NHE activity or the ability of Empa and Cariporide to inhibit NHE in isolated mouse cardiomyocytes. NHE activity as shown for pH recovery following NH4+ pulse in presence (A) and absence (B) of insulin. Both Cariporide and Empa inhibited NHE. N = 8 cardiomyocytes/group from five mice. ***P < 0.001 by one-way ANOVA with Dunnett’s post hoc vs. vehicle. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.

Figure 6 Increasing Empa dosages further delayed TOC, but did not result in reduced IR injury (as compared to ... Figure 6 Increasing Empa dosages further delayed TOC, but did not result in reduced IR injury (as compared to Figure 3). Hearts were subjected to IR and 3 µM Empa, 10 µM Empa or vehicle in the absence of insulin (A). Contracture development during ischaemia (B), TOC (C), global infarct size (D), RPP recovery at t = 2 h reperfusion relative to baseline RPP (E), end-diastolic pressure (F), and total LDH release during reperfusion (G) after 2 h reperfusion. N = 5 for vehicle, n = 5 for 3 µM Empa, and n = 6 for 10 µM Empa. Scale bar = 0.5 mm. LDH AUC, Lactate dehydrogenase activity area under the curve during reperfusion, normalized to heart wet weight; Pdia, diastolic pressure; RPP, rate pressure product; TOC, time of onset contracture. Unless provided in the caption above, the following copyright applies to the content of this slide: Published on behalf of the European Society of Cardiology. All rights reserved. © The Author(s) 2019. For permissions, please email: journals.permissions@oup.com.This article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Cardiovasc Res, cvz004, https://doi.org/10.1093/cvr/cvz004 The content of this slide may be subject to copyright: please see the slide notes for details.