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In vitro Assessment of Myocardial Protection following Hypothermia-Preconditioning in a Human Cardiac Myocytes Model

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Endothelial Cell Apoptosis but Not Necrosis Is Inhibited by Ischemic Preconditioning.

Jarosław Zalewski¹, Marta Szajna², Konrad Stępień^1,3

¹Department of Coronary Artery Disease and Heart Failure, Jagiellonian University Medical College, 31-202 Krakow, Poland.

International Journal of Molecular Sciences

|January 27, 2024

Summary

Ischemic preconditioning (IP) significantly inhibits apoptosis in endothelial cells (ECs) subjected to hypoxia/reoxygenation (HR). While IP delays necrosis, it effectively reduces both membrane and nuclear apoptosis, altering the Bcl-2/Bax ratio.

Keywords:

apoptosis endothelium hypoxia ischemic preconditioning reoxygenation

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Published on: May 9, 2013

Area of Science:

Cardiovascular Biology
Cellular Physiology
Molecular Medicine

Background:

Endothelial cells (ECs) are crucial for vascular function.
Hypoxia/reoxygenation (HR) induces EC death, contributing to various pathologies.
Ischemic preconditioning (IP) is a phenomenon that protects tissues from ischemic injury.

Purpose of the Study:

To investigate the protective effects of ischemic preconditioning (IP) against hypoxia/reoxygenation (HR)-induced endothelial cell (EC) death.
To quantify the impact of IP on both necrosis and apoptosis in ECs.
To analyze the molecular mechanisms, including Bcl-2/Bax gene expression, underlying IP's protective effects.

Main Methods:

Human umbilical vein endothelial cells (HUVECs) were exposed to hypoxia (2 or 6 hours) followed by reoxygenation.
IP was induced via a brief period of hypoxia and reoxygenation.
Cell death was assessed using lactate dehydrogenase (LDH) release for necrosis and propidium iodide/annexin V (PI/AV) staining, TUNEL assay, and RT-PCR for apoptosis and gene expression (Bcl-2, Bax).

Main Results:

HR induced significant necrosis (30-33%) and apoptosis (16-21%) in ECs.
IP reduced membrane apoptosis by 50% and nuclear apoptosis by 55-60% compared to HR.
IP treatment led to a significantly higher Bcl-2/Bax ratio, indicating an anti-apoptotic effect, which was modulated by hypoxia duration.

Conclusions:

Ischemic preconditioning (IP) offers significant protection against hypoxia/reoxygenation (HR)-induced endothelial cell (EC) apoptosis.
IP delays but does not prevent EC necrosis.
The protective mechanism involves an increased Bcl-2/Bax ratio, suggesting modulation of intrinsic apoptotic pathways.