Objective To investigate the role of hesperidin（Hsd）in attenuating hypoxic injury in H9c2 cells. Methods H9c2 cells divided into three groups: the normoxia group, the hypoxia group, and the hypoxia + Hsd group (H+Hsd). In addition, a hypoxic +Hsd+JNK agonist anisomycin group (designated as H+Hsd+Ani) was established to investigate the mechanism underlying the action of Hsd on H9c2 cells. The normoxia group group was maintained at 37°C with 5% CO2. The H + Hsd group received pre-treatment with varying concentrations of Hsd (5, 10, 15, 20, 40, 50, and 80 µmol/L) under normoxic conditions for 12 hours before being transferred to a hypoxic workstation for exposure to low oxygen levels (0.5% O2) for a duration of 48 hours. Cell viability was assessed using the CCK8 assay. Apoptosis and reactive oxygen species levels were evaluated by flow cytometry. The content of malondialdehyde (MDA) and superoxide dismutase (SOD) activity within cells was measured using corresponding kits. Creatine kinase MB isoenzyme (CKMB) and cardiac troponin I (cTn-I) levels were quantified via enzyme-linked immunosorbent assay (ELISA). Lactate dehydrogenase (LDH) activity was determined using an LDH assay kit. Corresponding protein levels were analyzed by western blotting techniques, while cellular immunofluorescence assay was employed to detect p65 nuclear translocation. Results Compared to the normoxia group, the hypoxia group exhibited reduced proliferation capacity (P<0.05), increased apoptosis rate and oxidative stress (P<0.01). Additionally, phosphorylation levels of JNK, JUN, and p65 were elevated (P<0.05), nucleation of p65 was enhanced. Meanwhile, the degradation of IKB alpha was observed (P<0.05). Conclusion Hsd has the potential to protect H9c2 cells against the damages induced by hypoxia. The underlying mechanism is associated with the JNK-JUN-NF-kB signaling pathway, by reducing the rate of apoptosis, mitigating oxidative stress.
Key words
hesperidin /
hypoxia /
myocardial cell /
myocardial damage
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