Affiliations: Department of Traditional Chinese Medicine and Medical History Literature, School of Basic Medicine, Shijiazhuang, Hebei 050200, P.R. China, Department of Traditional Chinese Medicine, School of Pharmacy, Hebei University of Chinese Medicine, Shijiazhuang, Hebei 050200, P.R. China, Department of Pharmacognosy, School of Pharmacy, Hebei Medical University, Shijiazhuang, Hebei 050200, P.R. China, Department of Diagnostics, Hebei Key Laboratory of Integrative Medicine on Liver‑Kidney Patterns, Institute of Integrative Medicine, College of Integrative Medicine, Hebei University of Chinese Medicine, Shijiazhuang, Hebei 050200, P.R. China, Department of Scientific Research Management, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China, Department of Hospital Management and Medical History Literature, Hebei Province Hospital of Chinese Medicine, The First Affiliated Hospital, Hebei University of Chinese Medicine, Shijiazhuang, Hebei 050200, P.R. China
Published online on:October 1, 2021https://doi.org/10.3892/etm.2021.10836
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 CoCl2 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 Ca2+ 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 Ca2+ 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.
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