Silencing of TXNIP attenuates oxidative stress injury in HEI-OC1 by inhibiting the activation of NLRP3 and NF-κB

Abstract

Sensorineural hearing loss (SNHL) is the most common type of hearing loss worldwide. The primary mechanism is oxidative injury to the cochlea as a result of oxidative stress. Therefore, exploring antioxidant strategies is particularly important in addressing SNHL.Thioredoxin-interacting protein (TXNIP) is an upstream target of oxidative stress-induced damage, and the NOD-like receptor protein 3 (NLRP3) and NF-κB pathways may be the main downstream molecular pathways, but this has not been reported in SNHL. Therefore, we investigated the molecular mechanism and role of TXNIP in oxidative stress injury induced by H2O2 in the HEI-OC1 auditory cells. To induce oxidative stress, HEI-OC1 cells were treated with H2O2. The TXNIP expression was measured by western blotting and Immunofluorescence. Intracellular TXNIP was knocked down using small interfering RNAs (siRNAs). Cell viability was measured by CCK8, total intracellular reactive oxygen species (ROS) by DCFH-DA, mitochondrial ROS by Mito-SOX, NLRP3, pro-caspase-1, total p65 NF-κB, and phospho-p65 NF-κB expression were measured by western blotting. Statistical analyses were performed using one-way analysis of variance, and p < 0.05 was considered statistically significant. We found that H2O2 treatment induced oxidative stress injury in HEI-OC1 cells, as evidenced by decreased cell viability and increased total intracellular and mitochondrial ROS levels (p < 0.05). TXNIP expression was elevated, and NLRP3 and NF-κB were activated (p < 0.05). Moreover, siRNA-TXINIP co-treatment reversed these changes and protected HEI-OC1 cells from oxidative stress (p < 0.05). We concluded that H2O2-induced oxidative stress in HEI-OC1 cells was alleviated by TXNIP inhibition. The finding may provide new insight into the prevention and treatment of SNHL.