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Catestatin improves post-ischemic left ventricular function and decreases ischemia/reperfusion injury in heart.

Penna C, Alloatti G, Gallo MP, Cerra MC, Levi R, Tullio F, Bassino E, Dolgetta S, Mahata SK, Tota B, Pagliaro P - Cell. Mol. Neurobiol. (2010)

Bottom Line: PostC reduced infarct size to 34 ± 5%.CST-Post reduced post-ischemic rise of diastolic LVP, an index of contracture, and significantly improved post-ischemic recovery of developed LVP.These results suggest a novel cardioprotective role for CST, which appears mainly due to a direct reduction of post-ischemic myocardial damages and dysfunction, rather than to an involvement of adrenergic terminals and/or endothelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Turin, Turin, Italy.

ABSTRACT
The Chromogranin A (CgA)-derived anti-hypertensive peptide catestatin (CST) antagonizes catecholamine secretion, and is a negative myocardial inotrope acting via a nitric oxide-dependent mechanism. It is not known whether CST contributes to ischemia/reperfusion injury or is a component of a cardioprotective response to limit injury. Here, we tested whether CST by virtue of its negative inotropic activity improves post-ischemic cardiac function and cardiomyocyte survival. Three groups of isolated perfused hearts from adult Wistar rats underwent 30-min ischemia and 120-min reperfusion (I/R, Group 1), or were post-conditioned by brief ischemic episodes (PostC, 5-cycles of 10-s I/R at the beginning of 120-min reperfusion, Group 2), or with exogenous CST (75 nM for 20 min, CST-Post, Group-3) at the onset of reperfusion. Perfusion pressure and left ventricular pressure (LVP) were monitored. Infarct size was evaluated with nitroblue-tetrazolium staining. The CST (5 nM) effects were also tested in simulated ischemia/reperfusion experiments on cardiomyocytes isolated from young-adult rats, evaluating cell survival with propidium iodide labeling. Infarct size was 61 ± 6% of risk area in hearts subjected to I/R only. PostC reduced infarct size to 34 ± 5%. Infarct size in CST-Post was 36 ± 3% of risk area (P < 0.05 respect to I/R). CST-Post reduced post-ischemic rise of diastolic LVP, an index of contracture, and significantly improved post-ischemic recovery of developed LVP. In isolated cardiomyocytes, CST increased the cell viability rate by about 65% after simulated ischemia/reperfusion. These results suggest a novel cardioprotective role for CST, which appears mainly due to a direct reduction of post-ischemic myocardial damages and dysfunction, rather than to an involvement of adrenergic terminals and/or endothelium.

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Cardiomyocytes survival after simulated I/R: representative experiment of a simulated I/R experiment: freshly isolated cardiomyocytes adhered on glass bottom dishes were placed under the confocal microscope and processed with the in vitro ischemia/reperfusion protocol as indicated in Fig. 1b. Cell viability was assessed by monitoring the time course of propidium iodide (PI) staining. Confocal Image acquisitions for each experimental condition (Tyr, IB, or CST) were performed at the times I, II, and III indicated Fig. 1b. Cells damaged by I/R are indicated by white (pre-reperfusion) or black (post-reperfusion) arrows (for further explanation see text). Bar graph summarizes the viability rate in the control protocol (Ctrl: 81.25%, 28 cells from three different experiments), in the I/R protocol (IB: 64 cells from seven experiments), and in the catestatin protocol (IB + CST: 45 cells from six experiments). Viability rate was quantified as the number of unstained cells at the end of reperfusion (PostRep) with respect to the number of unstained cells at t = 0 (PreRep) (no cells PostRep/no cells PreRep) × 100. Results are presented as mean ± SEM. * P < 0.05
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Fig4: Cardiomyocytes survival after simulated I/R: representative experiment of a simulated I/R experiment: freshly isolated cardiomyocytes adhered on glass bottom dishes were placed under the confocal microscope and processed with the in vitro ischemia/reperfusion protocol as indicated in Fig. 1b. Cell viability was assessed by monitoring the time course of propidium iodide (PI) staining. Confocal Image acquisitions for each experimental condition (Tyr, IB, or CST) were performed at the times I, II, and III indicated Fig. 1b. Cells damaged by I/R are indicated by white (pre-reperfusion) or black (post-reperfusion) arrows (for further explanation see text). Bar graph summarizes the viability rate in the control protocol (Ctrl: 81.25%, 28 cells from three different experiments), in the I/R protocol (IB: 64 cells from seven experiments), and in the catestatin protocol (IB + CST: 45 cells from six experiments). Viability rate was quantified as the number of unstained cells at the end of reperfusion (PostRep) with respect to the number of unstained cells at t = 0 (PreRep) (no cells PostRep/no cells PreRep) × 100. Results are presented as mean ± SEM. * P < 0.05

Mentions: The protective role of CST was also investigated on isolated adult cardiomyocytes subjected to simulated I/R, as indicated in the “Methods” (Fig. 1b). Image acquisitions for each experimental condition (Ctrl, IB, or IB + CST) were performed at the times I, II, and III, showed in Fig. 1b. Representative experiments are presented in Fig. 4a. With respect to control (Ctrl), the appearance of propidium iodide staining at the end of reperfusion (III) in the ischemic (IB) sample clearly demonstrates the effectiveness of I/R simulation. CST (5 nM) administration (IB + CST) significantly (P < 0.05, Fig. 4b) preserved cell viability after reperfusion, being comparable to that observed in control condition. Figure 4b summarizes the results of these experiments: viability rate was 81.3 ± 10.8% in control, 12.9 ± 8.3% in IB, and 63.5 ± 17.0% in IB + CST.Fig. 4


Catestatin improves post-ischemic left ventricular function and decreases ischemia/reperfusion injury in heart.

Penna C, Alloatti G, Gallo MP, Cerra MC, Levi R, Tullio F, Bassino E, Dolgetta S, Mahata SK, Tota B, Pagliaro P - Cell. Mol. Neurobiol. (2010)

Cardiomyocytes survival after simulated I/R: representative experiment of a simulated I/R experiment: freshly isolated cardiomyocytes adhered on glass bottom dishes were placed under the confocal microscope and processed with the in vitro ischemia/reperfusion protocol as indicated in Fig. 1b. Cell viability was assessed by monitoring the time course of propidium iodide (PI) staining. Confocal Image acquisitions for each experimental condition (Tyr, IB, or CST) were performed at the times I, II, and III indicated Fig. 1b. Cells damaged by I/R are indicated by white (pre-reperfusion) or black (post-reperfusion) arrows (for further explanation see text). Bar graph summarizes the viability rate in the control protocol (Ctrl: 81.25%, 28 cells from three different experiments), in the I/R protocol (IB: 64 cells from seven experiments), and in the catestatin protocol (IB + CST: 45 cells from six experiments). Viability rate was quantified as the number of unstained cells at the end of reperfusion (PostRep) with respect to the number of unstained cells at t = 0 (PreRep) (no cells PostRep/no cells PreRep) × 100. Results are presented as mean ± SEM. * P < 0.05
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Fig4: Cardiomyocytes survival after simulated I/R: representative experiment of a simulated I/R experiment: freshly isolated cardiomyocytes adhered on glass bottom dishes were placed under the confocal microscope and processed with the in vitro ischemia/reperfusion protocol as indicated in Fig. 1b. Cell viability was assessed by monitoring the time course of propidium iodide (PI) staining. Confocal Image acquisitions for each experimental condition (Tyr, IB, or CST) were performed at the times I, II, and III indicated Fig. 1b. Cells damaged by I/R are indicated by white (pre-reperfusion) or black (post-reperfusion) arrows (for further explanation see text). Bar graph summarizes the viability rate in the control protocol (Ctrl: 81.25%, 28 cells from three different experiments), in the I/R protocol (IB: 64 cells from seven experiments), and in the catestatin protocol (IB + CST: 45 cells from six experiments). Viability rate was quantified as the number of unstained cells at the end of reperfusion (PostRep) with respect to the number of unstained cells at t = 0 (PreRep) (no cells PostRep/no cells PreRep) × 100. Results are presented as mean ± SEM. * P < 0.05
Mentions: The protective role of CST was also investigated on isolated adult cardiomyocytes subjected to simulated I/R, as indicated in the “Methods” (Fig. 1b). Image acquisitions for each experimental condition (Ctrl, IB, or IB + CST) were performed at the times I, II, and III, showed in Fig. 1b. Representative experiments are presented in Fig. 4a. With respect to control (Ctrl), the appearance of propidium iodide staining at the end of reperfusion (III) in the ischemic (IB) sample clearly demonstrates the effectiveness of I/R simulation. CST (5 nM) administration (IB + CST) significantly (P < 0.05, Fig. 4b) preserved cell viability after reperfusion, being comparable to that observed in control condition. Figure 4b summarizes the results of these experiments: viability rate was 81.3 ± 10.8% in control, 12.9 ± 8.3% in IB, and 63.5 ± 17.0% in IB + CST.Fig. 4

Bottom Line: PostC reduced infarct size to 34 ± 5%.CST-Post reduced post-ischemic rise of diastolic LVP, an index of contracture, and significantly improved post-ischemic recovery of developed LVP.These results suggest a novel cardioprotective role for CST, which appears mainly due to a direct reduction of post-ischemic myocardial damages and dysfunction, rather than to an involvement of adrenergic terminals and/or endothelium.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical and Biological Sciences, University of Turin, Turin, Italy.

ABSTRACT
The Chromogranin A (CgA)-derived anti-hypertensive peptide catestatin (CST) antagonizes catecholamine secretion, and is a negative myocardial inotrope acting via a nitric oxide-dependent mechanism. It is not known whether CST contributes to ischemia/reperfusion injury or is a component of a cardioprotective response to limit injury. Here, we tested whether CST by virtue of its negative inotropic activity improves post-ischemic cardiac function and cardiomyocyte survival. Three groups of isolated perfused hearts from adult Wistar rats underwent 30-min ischemia and 120-min reperfusion (I/R, Group 1), or were post-conditioned by brief ischemic episodes (PostC, 5-cycles of 10-s I/R at the beginning of 120-min reperfusion, Group 2), or with exogenous CST (75 nM for 20 min, CST-Post, Group-3) at the onset of reperfusion. Perfusion pressure and left ventricular pressure (LVP) were monitored. Infarct size was evaluated with nitroblue-tetrazolium staining. The CST (5 nM) effects were also tested in simulated ischemia/reperfusion experiments on cardiomyocytes isolated from young-adult rats, evaluating cell survival with propidium iodide labeling. Infarct size was 61 ± 6% of risk area in hearts subjected to I/R only. PostC reduced infarct size to 34 ± 5%. Infarct size in CST-Post was 36 ± 3% of risk area (P < 0.05 respect to I/R). CST-Post reduced post-ischemic rise of diastolic LVP, an index of contracture, and significantly improved post-ischemic recovery of developed LVP. In isolated cardiomyocytes, CST increased the cell viability rate by about 65% after simulated ischemia/reperfusion. These results suggest a novel cardioprotective role for CST, which appears mainly due to a direct reduction of post-ischemic myocardial damages and dysfunction, rather than to an involvement of adrenergic terminals and/or endothelium.

Show MeSH
Related in: MedlinePlus