Nicotinamide adenine dinucleotide inhibits the production of reactive oxygen species and myocardial cell pyroptosis caused by hypoxia/re-oxygenation injury

  • World J Cardiol. 2026 Jan 26;18(1):114108. doi: 10.4330/wjc.v18.i1.114108.
Shuang Dong  1 ,  Yun-Qi Liu  2 ,  Yi-Jun Tu  3 ,  Shan Gao  2 ,  Yu-Jie Liu  2 ,  Chang Liu  4 ,  Zuo-Wei Pei  1
Affiliations
  • 1. Department of Cardiology, Central Hospital of Dalian University of Technology, Dalian 116033, Liaoning Province, China.
  • 2. Central Laboratory, Central Hospital of Dalian University of Technology, Dalian 116033, Liaoning Province, China.
  • 3. Department of Pharmacy, Central Hospital of Dalian University of Technology, Dalian 116033, Liaoning Province, China.
  • 4. Central Laboratory, Central Hospital of Dalian University of Technology, Dalian 116033, Liaoning Province, China. [email protected].
PMID: 41607618 DOI: 10.4330/wjc.v18.i1.114108
Abstract

Background: Myocardial ischemia/reperfusion (I/R) is a significant factor that negatively impacts the treatment outcomes of coronary heart disease, particularly acute myocardial infarction. The oxidized form of nicotinamide adenine dinucleotide (NAD) - NAD+ is crucial for various cellular functions.

Aim: To explore the effects and mechanisms of NAD+ on cell death caused by hypoxia/re-oxygenation (H/R) injury in H9c2 cells.

Methods: Cell viability was assessed using the Cell Counting Kit-8 assay. Apoptosis in H9c2 cells was evaluated through terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling staining. Intracellular Reactive Oxygen Species levels were measured with the fluorescent probe dichloro-dihydrofluorescein diacetate. Intracellular NAD+ levels were quantified using a NAD/NAD reduced form assay kit. The impact of NAD+ on the expression of NOD-like Receptor pyrin domain-containing 3, apoptosis-associated speck-like protein containing a CARD, and Caspase-1 was analyzed by reverse transcription polymerase chain reaction and western blotting.

Results: The study demonstrated that NAD+ supplementation protects H9c2 cells from H/R induced cell Pyroptosis. Mechanistically, external NAD+ reduces H/R induced Pyroptosis in H9c2 cells by inhibiting the NOD-like Receptor pyrin domain-containing 3 inflammasome.

Conclusion: These results indicate that NAD+ supplementation may serve as a promising therapeutic strategy for I/R injury.

Keywords
Hypoxia/re-oxygenation; Ischemia/reperfusion injury; Nicotinamide adenine dinucleotide; Oxidative stress; Pyroptosis.
Products