Targeting necroptosis as therapeutic potential in chronic myocardial infarction

Table 2 Markers used for differentiation between apoptosis and necroptosis in the experimental settings

	Apoptosis	Necroptosis	References
Morphology	Cytoplasmic shrinkage Chromatin condensation (pyknosis) Nuclear fragmentation (karyorrhexis) Plasma membrane blebbing (zeiosis) Shedding of apoptotic bodies	Increasingly translucent cytoplasm Swelling of organelles Membrane permeabilization Increased cell volume (oncosis) Mild chromatin condensation (nuclei remain intact)	[11, 13, 20, 71, 73, 87]
Death execution events	Caspase-3 execution pathway causes cell shrinkage, chromosomal condensation and DNA fragmentation	MLKL phosphorylation and translocation to the plasma membrane causes membrane permeabilization	[11,12,13, 20, 27, 71, 73, 87, 88]
Death regulatory factors	BID, BAX, Bcl-2, Cytochrome c APAF1, FADD, Caspase-8, Caspase-9	RIPK1, RIPK3	[11,12,13, 20, 27, 58, 71, 73, 87, 88]
Death execution factors	Caspase-3 Caspase-7	p-MLKL	[11,12,13, 20, 27, 58, 71, 73, 87, 88]
Methods for evaluation in cardiomyocytes	TUNEL assays DNA laddering Annexin V positive Caspase-3/7 activity assay	PI staining (staining with impermeant dyes) HMGB1 release LDH assay Detection of RIPK1, RIPK3 and MLKL	[71, 87]

APAF1, Apoptotic protease activating factor 1; Bax, BCL2 associated X protein; BCL-2, B-cell lymphoma 2 protein; BID, BH3-interacting domain death agonist; FADD, Fas-associated protein with death domain; HMGB1, high mobility group box 1; LDH, Lactate dehydrogenase; MLKL, Mixed lineage kinase domain like pseudokinase; p-MLKL, the phosphorylated form of mixed lineage kinase domain like pseudokinase; RIPK1, receptor-interacting serine/threonine-protein kinase 1; RIPK3, receptor-interacting serine/threonine-protein kinase 3; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling

ISSN: 1423-0127