Objective To investigate the protective effect of puerarin (Pur) against hypoxia-induced pyroptosis and oxidative stress in H9c2 cardiomyocytes. Methods An in vitro model of hypoxia injury was established using H9c2 cardiomyocytes cultured in a hypoxic workstation (0.5% O₂). Cells were divided into five groups: normoxic control (normoxia), hypoxia (hypoxia), and hypoxia with Pur treatment at concentrations of 1 µmol/L (h+1Pur), 2.5 µmol/L (h+2.5Pur), and 5 µmol/L (h+5Pur), each with three replicates. After a 12-hour pre-treatment with Pur, the hypoxia groups were subjected to hypoxia for 48 hours. Cell viability, intracellular reactive oxygen species (ROS), and supernatant levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6), were measured. Additionally, the expression of pyroptosis-related proteins (NLRP3, Caspase-1, GSDMD-N, and IL-18) was assessed by Western blot. Results Compared with the normoxic control, hypoxia significantly reduced cell viability and SOD activity (P<0.05), and markedly increased intracellular ROS, LDH, and MDA levels (P<0.05). Moreover, the protein levels of NLRP3, Caspase-1, and GSDMD-N were significantly up-regulated (P<0.05). Pur treatment significantly improved cell viability and SOD activity, and reduced ROS, LDH, and MDA levels (P<0.05). Importantly, Pur markedly down-regulated the expression of NLRP3, GSDMD-N, and IL-18 (P<0.05) compared with the hypoxia group. Conclusion Puerarin effectively attenuates hypoxia-induced pyroptosis and oxidative stress in H9c2 cells, partly through inhibition of the NLRP3/Caspase-1/GSDMD signaling pathway.
Key words
puerarin /
cardiomyocytes /
hypoxia /
pyroptosis /
oxidative stress
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