Effect of lycopene preconditioning on TLRs/NF-κB signaling pathway during hypoxia-reoxygenation injury to rat cardiomyocytes / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the effect of lycopene preconditioning on Toll-like receptors (TLRs)/nuclear factor kappa B (NF-κB) signaling pathway during hypoxia-reoxygenation (H/R) injury to rat cardiomyocytes.Methods:The rat H9C2 cardiomyocytes were cultured in vitro, inoculated in a petri dish at a density of 8×10 4 cells/ml and divided into 3 groups( n=30 each) using a random number table method: control group (C group), H/R group and lycopene preconditioning group (LP group). The H9C2 cardiomyocytes were reoxygenated for 6 h after hypoxia to establish a model of H/R injury in H/R group and LP group.The cells were preconditioned with lycopene 20 μmol/L at 12 h before establishing the model in LP group.The cell viability was detected by CCK-8 assay.The cell apoptosis rate was detected by flow cytometry (Annexin V and PI double staining). The levels of lactic dehydrogenase (LDH), superoxide dismutase (SOD) and malondialdehyde (MDA) and reactive oxygen species (ROS) were detected by microplate method.The expression of TLR2, TLR3 and NF-κB was detected by Western blot. Results:Compared with group C, the cell viability and SOD level were significantly decreased, the apoptosis rate and levels of LDH, MDA and ROS were increased, and the expression of TLR2, TLR3 and NF-κB was up-regulated in H/R and LP groups ( P<0.05). Compared with group H/R, the cell viability and SOD level were significantly increased, the apoptosis rate and levels of LDH, MDA and ROS were decreased, and the expression of TLR2, TLR3 and NF-κB was down-regulated in group LP ( P<0.05). Conclusion:The mechanism by which lycopene preconditioning attenuates H/R injury may be related to inhibiting activation of TLRs/NF-κB signaling pathway and inhibiting oxidative stress response in rat cardiomyocytes.