Protective effects of safranal on hypoxia/reoxygenation‑induced injury in H9c2 cardiac myoblasts via the PI3K/AKT/GSK3β signaling pathway

Abstract

Safranal (SFR), an active ingredient extracted from saffron, exhibits a protective effect on the cardiovascular system. However, the mechanism of SFR against hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury has previously not been investigated in vitro. The aim of the present study was therefore to observe the protective effects of SFR on H/R-induced cardiomyocyte injury and to explore its mechanisms. A H/R injury model of H9c2 cardiac myoblasts was established by administering 800 µmol/l CoCl 2 to H9c2 cells for 24 h and reoxygenating the cells for 4 h to induce hypoxia. H9c2 cardiac myoblasts were pretreated with SFR for 12 h to evaluate the associated protective effects. A Cell Counting Kit-8 assay was used for cell viability detection, and the expression levels of lactate dehydrogenase (LDH), creatine kinase-MB (CK-MB), glutathione peroxidase (GSH-px), catalase (CAT), superoxide dismutase (SOD), malondialdehyde (MDA) and caspase-3, and the intracellular Ca 2+ concentration were measured using the corresponding commercial kits. Levels of reactive oxygen species (ROS) in the cells were detected using 2,7-dichlorodihydrofluorescein diacetate. Flow cytometry was used to determine the degree of apoptosis and the level of mitochondrial membrane potential (MMP). Moreover, the expression levels of phosphorylated (p-)PI3K, AKT, p-AKT, glycogen synthase kinase 3β (GSK3β), p-GSK3β, Bcl-2, Bax, caspase-3 and cleaved caspase-3 were measured using western blot analysis. Results of the present study demonstrated that the H9c2 cardiac myoblasts treated with SFR exhibited significantly improved levels of viability and significantly reduced levels of ROS, compared with the H/R group. Furthermore, compared with the H/R group, SFR treatment significantly increased the MMP levels and antioxidant enzyme levels, including CAT, SOD and GSH-px; whereas the levels of CK-MB, LDH, MDA and intracellular Ca 2+ concentration were significantly decreased. Moreover, the results of the present study demonstrated that SFR significantly reduced caspase-3, cleaved caspase-3 and Bax protein expression levels, but upregulated the Bcl-2 protein expression levels. SFR also increased the protein expressions of PI3K/AKT/GSK3β. In summary, the results suggested that SFR may exert a protective effect against H/R-induced cardiomyocyte injury, which occurs in connection with the inhibition of oxidative stress and apoptosis via regulation of the PI3K/AKT/GSK3β signaling pathway.

Tanshinone IIA confers protection against myocardial ischemia/reperfusion injury by inhibiting ferroptosis and apoptosis via VDAC1

“…Such injury is known as IRI ( 41 ). Previous studies have reported that the pathogenesis of IRI involves excessive ROS generation and inflammatory responses ( 42 , 43 ). The underlying molecular mechanisms and pathways of IRI are complicated and associated with multiple forms of regulated cell death, including pyroptosis, necroptosis, ferroptosis and apoptosis ( 44 , 45 ).…”

Traditional chinese medicine in coronary microvascular disease

“…In addition, safranal, an active ingredient extracted from saffron, exerts a protective effect on the cardiovascular system. Wang H. et al (2021 ) reported that safranal could increase the viability of H9c2 cardiac myoblasts and alleviate H/R-induced H9c2 cardiac myoblast injuries via the PI3K/AKT/GSK3β signaling pathway.…”

Safranal alleviates pressure overload induced-cardiac hypertrophy and dysfunction via activating AMPK signaling pathway

“…The latest research showed that SFR had some protective effect on the heart. For example, Wang H, et al thought that SFR exerted protective effect against hypoxia/reoxygenation -induced cardiomyocyte injury via inhibiting oxidative stress and apoptosis [10]. Mehdizadeh R, et al, revealed that SFR played cardio-protective on myocardial infarction by inhibitting of oxidative stress and regulating Ca2+ homeostasis [11,12].…”