Exogenous ARC down-regulates caspase-3 expression and inhibits apoptosis of broiler chicken cardiomyocytes exposed to hydrogen peroxide

Abstract

Apoptosis repressor with caspase recruitment domain (ARC) is highly involved in apoptosis induced by oxidative stress or ischaemia/reperfusion injury. Furthermore, even though the exact mechanism is still unknown, some studies suggest that exogenous ARC also possesses anti-apoptotic ability. The study investigated whether mouse-derived ARC acquires anti-apoptotic ability and the pathway of regulation in chick embryo cardiomyocytes. To evaluate whether mouse-derived ARC can inhibit chick embryo cardiomyocyte apoptosis induced by hydrogen peroxide, recombinant pcDNA3.1/ARC plasmid was acquired and transfected into chick embryo cardiomyocytes. ARC-related gene (caspase-2, caspase-8, caspase-3, and caspase-9, cytochrome C, bcl-2, and XIAP) mRNA and protein expression levels were detected by real-time polymerase chain reaction and western blotting, respectively. Here we demonstrate that hydrogen peroxide induced apoptosis in chick embryo cardiomyocytes in a time-dependent manner and that this effect could be suppressed by mouse-derived ARC expression. Moreover, unlike endogenous ARC, exogenous ARC was exclusively expressed in the cytoplasm and down-regulated caspase-2, caspase-8, and caspase-3, bcl-2, and XIAP gene expression levels. However, only caspase-3 protein levels were decreased. In addition, threonine 149 phosphorylation by CK2 was required for exogenous ARC to exert an antiapoptotic effect in chicken embryo cardiomyocytes and suggested exogenous ARC may in part share the same pathway of regulation with endogenous ARC. These results indicate that mouse-derived ARC plays an important role in protection of chick embryo cardiomyocytes against oxidative stress apoptosis by inhibiting caspase-3 mRNA and protein expression levels.

Effect of metoprolol on myocardial apoptosis after coronary microembolization in rats

“…[20] Caspase-3 mediates the pathway to apoptosis protease cascade, as the key protease to mammalian cell apoptosis, the main apoptosis effect factor and the most important apoptosis performer. [21,22] Intravenous injection of metoprolol before surgery could significantly improve the myocardial apoptosis and systolic dysfunction caused by CME. Molecular biology showed the significantly inhibited caspase-3 protein activation in myocardial tissue, and reduced myocardial apoptosis rate.…”

Postconditioning signalling in the heart: mechanisms and translatability

“…Mechanical postconditioning has been shown to decrease TNF‐α and limit ROS formation during early reperfusion, resulting in attenuation of apoptosis (Kin et al ., ). Most recently, the apoptosis repressor with caspase recruitment domain has been shown to decrease caspase‐3 activity and subsequent apoptosis in chick embryo myocytes following exposure to hydrogen peroxide (Wu et al ., ).…”

Stanniocalcin-1 protects bovine intestinal epithelial cells from oxidative stress-induced damage

“…Therefore, cultured primary IECs were harvested and used for our investigation. To mimic cellular damage induced by chronic enteritis, the primary cells were treated with H 2 O 2 for 4 ~ 24 h. This strategy helped ensure the continuity of reactive oxygen species (ROS) production found during chronic inflammation, and was based on previous work by Bae et al [ 2 ] and our group [ 28 ] who treated MCF-7 cells or cardiomyocytes for 24 h with 200 µM H2 2 O 2 to induce apoptosis.…”