Table 6.

Evidence of interventions to modify pyroptosis in MI: Reports from in vitro studies.

Condition/Hypoxia (h)	Model	Intervention	Pyroptotic marker				Inflammatory marker/ROS	Cell viability/Toxicity	Other cell death marker/relevant finding	Interpretation	Ref
			GSDMD		Caspase-1	Others
			Active form	Full form
2	H9C2 cells	Melatonin 10 μmol/L, during hypoxia	↓	–	↓	↓ NLRP3	↓ TLR4 ↓ NF-κB ↓ IL-1β ↓ IL-18	↓ LDH	–	Melatonin reduced pyroptosis through the canonical pathway.	42, 43
4	NRCMs	H2-containing medium, >0.6 mmol/L, during hypoxia	↓ GSDMD-N per GSDMD-FL ratio	↓		↓ ASC ↓ NLRP3	↓ IL-1β	↑ Cell viability ↓ LDH	↓ PI	H2 and MCC950 reduced pyroptosis through the canonical pathway. However, combined treatment did not provide further protective effect.	18
		MCC950 (NLRP3 inhibitor) 10 μmol/L, during hypoxia	↓ GSDMD-N per GSDMD-FL ratio	↓	↓ ASC ↓ NLRP3	↓ IL-1β	↑ Cell viability ↓ LDH	↓ PI
		H2 -containing medium + MCC950, during hypoxia	↓ GSDMD-N per GSDMD-FL ratio	↓	↓ ASC ↓ NLRP3	↓ IL-1β	↑ Cell viability ↓ LDH	↓ PI
6	H9C2 cells	hMSC-derived exosomes	↓	↓	↓	↓ ASC ↓ NLRP3	↓ IL-1β ↓ IL-18	↑ Cell viability	↓ TUNEL	Exosomal lncRNA KLF3-AS1 derived from hMSCs reduced pyroptosis through inhibition of miR-138-5p, and promoted Sirt1, subsequently reduced GSDMD.	42, 43
		KLF3-AS1 overexpression hMSC-derived exosomes	↓↓	↓↓	↓↓	↓↓ ASC ↓↓ NLRP3	↓↓ IL-1β ↓↓ IL-18	↑↑ Cell viability	↓↓ TUNEL
		hMSC-derived exosomes + miR-138-5p inhibitor cell transfection	↓↓	↓↓	↓↓	↓↓ ASC ↓↓ NLRP3	↓↓ IL-1β ↓↓ IL-18	↑↑ Cell viability	↓↓ TUNEL
		hMSC-derived exosomes + Sh-Sirt1 cell transfection vs. Hypoxia + exosomes	↑	↑	↑	↑ ASC ↑ NLRP3	↑ IL-1β ↑ IL-18	↓ Cell viability	↑ TUNEL
24	NMCMs	Kanglexin 10 μmol/L, during hypoxia	↓	↓	↓	↓ NLRP3	↓ IL-1β ↓ IL-18	–	↓ TUNEL ↓ PI	Kanglexin reduced pyroptosis through the canonical pathway.	42, 43
12 (+TNF-α)	H9C2 cells	Liraglutide 100 nmol/L, pretreatment 1 h before hypoxia	↓	–	↓	↓ NLRP3	↓ NOX4 ↓ ROS	↑ Cell viability ↓ LDH	↑ SIRT1 ↓ PI	Liraglutide pretreatment reduced oxidative stress and promoted SIRT1, subsequently reduced pyroptosis through the canonical pathway.	42, 43
		liraglutide pretreatment + EX527 (SIRT1 inhibitor) 200 nmol/L, pretreatment 1 h before hypoxia vs. hypoxia + liraglutide	↑	–	↑	↑ NLRP3	↑ NOX4 ↑ ROS	↓ Cell viability ↑ LDH	↑ PI
12 (+TNF-α)	H9C2 cells	Rosuvastatin 20 μmol/L, pretreatment 2 h before hypoxia	↓ GSDMD-N per GSDMD-FL ratio	↓		↓ NLRP3	↓ ROS	↑ Cell viability ↓ LDH	↓ PI	Rosuvastatin and NAC pretreatment reduced oxidative stress, subsequently reduced pyroptosis through the canonical pathway.	43
		NAC 5 μmol/L, pretreatment 2 h before hypoxia	↓ GSDMD-N per GSDMD-FL ratio	↓	↓ NLRP3	–	↑ Cell viability ↓ LDH	↓ PI
OGD 36 h	H9C2 cells	NAC 50 μmol/L, after 36 h OGD	↓	↓	↓	↓ ASC ↓ NLRP3	↓ p-NF-κB ↓ ROS ↑ SOD	↓ LDH	–	NAC and PDTC reduced oxidative stress, subsequently reduced pyroptosis through the canonical pathway.	42, 43
		PDTC 25 μmol/L, after 36 h OGD	↓	↓	↓	↓ ASC ↓ NLRP3	↓ p-NF-κB	↓ LDH	–

ASC: apoptosis-associated speck-like protein containing a caspase recruitment domain; GSDMD: Gasdermin D; GSDMD-FL: full-length Gasdermin D; GSDMD-N: N-terminal Gasdermin D fragment; H: hypoxia; H/R: hypoxia/reoxygenation; hMSC: human mesenchymal stem cell; IL: interleukin; KLF3-AS1: KLF3 antisense RNA1; LDH: lactate dehydrogenase; lncRNA: long non-coding RNA; miR: microRNA; NAC: N-acetylcysteine; NF-κB: nuclear factor κ-light-chain-enhancer of activated B cells; NLRP3: NACHT, LRR and PYD domains-containing protein 3; NMCMs: neonatal mouse cardiomyocytes; NOX4: nicotinamide adenine dinucleotide phosphate oxidase 4; NRCMs: neonatal rat cardiomyocytes; OGD: oxygen-glucose deprivation; PDTC: pyrrolidine dithiocarbamate; PI: propidium iodide; R: reoxygenation; ROS: reactive oxygen species; sh-Sirt1: short hairpin Sirtuin 1; Sirt1: Sirtuin 1; SOD: superoxide dismutase; TNF-α: tumor necrosis factor alpha; TUNEL: terminal deoxynucleotidyl transferase dUTP nick end labeling.