Journal Article

Azelnidipine protects HL-1 cardiomyocytes from hypoxia/reoxygenation injury by enhancement of NO production independently of effects on gene expression

Minato, Hiroyuki, Otsuki, Akihiro, Yamamoto, Kazuhiro, Kinugasa, Yoshiharu, Hisatome, Ichiro, Wakimizu, Takayuki, Shirayoshi, Yasuaki, Tsuneto, Motokazu, Endo, Ryo, Ninomiya, Haruaki, Notsu, Tomomi, Kurata, Yasutaka

2024

Overview

It remains to be elucidated whether Ca 2+ antagonists induce pharmacological preconditioning to protect the heart against ischemia/reperfusion injury. The aim of this study was to determine whether and how pretreatment with a Ca 2+ antagonist, azelnidipine, could protect cardiomyocytes against hypoxia/reoxygenation (H/R) injury in vitro. Using HL-1 cardiomyocytes, we studied effects of azelnidipine on NO synthase (NOS) expression, NO production, cell death and apoptosis during H/R. Action potential durations (APDs) were determined by the whole-cell patch-clamp technique. Azelnidipine enhanced endothelial NOS phosphorylation and NO production in HL-1 cells under normoxia, which was abolished by a heat shock protein 90 inhibitor, geldanamycin, and an antioxidant, N -acetylcysteine. Pretreatment with azelnidipine reduced cell death and shortened APDs during H/R. These effects of azelnidipine were diminished by a NOS inhibitor, L-NAME, but were influenced by neither a T-type Ca 2+ channel inhibitor, NiCl 2 , nor a N-type Ca 2+ channel inhibitor, ω-conotoxin. The azelnidipine-induced reduction in cell death was not significantly enhanced by either additional azelnidipine treatment during H/R or increasing extracellular Ca 2+ concentrations. RNA sequence (RNA-seq) data indicated that azelnidipine-induced attenuation of cell death, which depended on enhanced NO production, did not involve any significant modifications of gene expression responsible for the NO/cGMP/PKG pathway. We conclude that pretreatment with azelnidipine protects HL-1 cardiomyocytes against H/R injury via NO-dependent APD shortening and L-type Ca 2+ channel blockade independently of effects on gene expression.

• Publisher: Springer Japan,Springer Nature B.V
• Subject: Acetylcysteine / Action potential / Action Potentials - drug effects / Animals / Apoptosis / Apoptosis - drug effects / Azetidinecarboxylic Acid - analogs & derivatives / Azetidinecarboxylic Acid - pharmacology / Biomedical Engineering and Bioengineering / Calcium (extracellular) / Calcium antagonists / Calcium Channel Blockers - pharmacology / Calcium channels (L-type) / Calcium channels (N-type) / Calcium channels (T-type) / Calcium ions / Cardiac Surgery / Cardiology / Cardiomyocytes / Cell death / Cell Hypoxia / Cell Line / Dihydropyridines - pharmacology / Geldanamycin / Gene expression / Gene Expression Regulation - drug effects / Heat shock proteins / Hsp90 protein / Hypoxia / Inhibitors / Injuries / Ischemia / Medicine / Medicine & Public Health / Mice / Mortality / Myocardial Reperfusion Injury - genetics / Myocardial Reperfusion Injury - metabolism / Myocardial Reperfusion Injury - pathology / Myocardial Reperfusion Injury - prevention & control / Myocytes, Cardiac - drug effects / Myocytes, Cardiac - metabolism / Myocytes, Cardiac - pathology / NG-Nitroarginine methyl ester / Nickel chloride / Nitric oxide / Nitric Oxide - metabolism / Nitric Oxide Synthase Type III - metabolism / Nitric-oxide synthase / Nucleotide sequence / Original Article / Phosphorylation / Preconditioning / Pretreatment / Proteins / Reperfusion / Ribonucleic acid / RNA / Software / Vascular Surgery