Tannic acid attenuates hypoxia-induced damage in H9c2 cardiomyocytes by inhibiting pyroptosis
Zheng Tao
Ma Long
Zhang Longjiang
Abstract：Objective To investigate the cytoprotective effect and mechanism of tannic acid(TA)against hypoxia-induced damage in H9c2 cardiomyocytes.Methods A model of hypoxia-induced cell injury was established in rat H9c2 cardiomyocytes with conditioned medium and a hypoxia chamber.H9c2 cells were divided into control group,hypoxia group 1,low-and high-dose TA groups(0.2 and 0.8 µmol TA).Functional recovery experiments were performed using rotenone(Rot),and the cells were divided into hypoxia group 2,hypoxia+Rot group(50 nmol Rot),hy-poxia+Rot+high-dose TA group(50 nmol Rot+0.8 μmol TA),hypoxia+high-dose TA group(0.8 μmol TA).Cellular damage was assessed using CCK-8 assay and LDH test.Intracellular lev-els of ROS and mitochondrial reactive oxygen species(mitoSOX),cell apoptosis,and protein ex-pression of NLRP3,Caspase-1,GSDMD,and IL-1β were detected by chemiluminescent assay,flow cytometry,and Western blotting,respectively.ELISA was employed to measure the secretion of IL-1β.Results Compared to the control group,the hypoxia group 1 exhibited notably reduced cell density and viability,increased LDH activity and cell apoptotic rate,and elevated protein levels of NLRP3,Cleaved-Caspase-1,Cleaved-GSDMD,and Cleaved-IL-1β(P＜0.05).Both low-and high-dose TA treatment improved cell viability,reduced LDH activity and apoptotic rate,and decreased the protein levels of NLRP3,Cleaved-Caspase-1,Cleaved-GSDMD,and Cleaved-IL-1β and the se-cretion of IL-1β when compared with the hypoxia group 1(P＜0.05).Significant increases were observed in intracellular mitoSOX levels and the expression of NLRP3 and Cleaved-IL-1β at pro-tein level in the hypoxia+Rot group compared to the hypoxia group 2(P＜0.05).Similarly,the hypoxia+Rot+high-dose TA group also exhibited significant increases in mitoSOX and NLRP3 and Cleaved-IL-1β levels when compared to the hypoxia+high-dose TA group(0.85±0.02 vs 0.40±0.03,0.61±0.03 vs 0.47±0.05,0.70±0.06 vs 0.48±0.09,P＜0.05).Conclusion TA alleviates hypoxia-induced H9c2 cell injury by inhibiting ROS/NLRP3 pathway-mediated pyroptosis.
Keywords：pyroptosishypoxiamitochondriarat H9c2 cardiomyocytestannic acid
Publication Date：2024-07-15
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 811-816 )