Hydrogen peroxide-mediated oxidative stress induces inflammasome activation in term human placental explants

The placenta is a multifunctional organ that can suffer with imbalances between pro- and antioxidant molecules, contributing for inflammatory imbalance. The inflammation generated by oxidative stress may induce inflammasome activation, an essential complex for pro-inflammatory cytokine production.

The aim of this study was to evaluate whether hydrogen peroxide (H₂O₂) mediated oxidative stress induces inflammasome activation on placental explants.

Tissue cultures of placental explants obtained from normotensive pregnant women were performed in different concentrations of H₂O₂. Gene expressions of NLRP3, caspase-1, IL-1β, TNF-α and IL-10 were evaluated by qPCR. Superoxide dismutase (SOD), catalase, Heat shock protein 70 (Hsp70), Caspase-1, TNF-α, IL-1β, IL-10 and human Chorionic Gonadotropin (hCG) were determined by ELISA.

Concentrations of catalase, Hsp70, hCG and SOD were higher in cultures with 100 and 1000 µM H₂O₂ compared to controls. Gene and protein expressions of TNF-α and IL-1β were elevated in cultures with 1000 μM H₂O₂ compared to controls. This concentration led to inflammasome activation, by increasing gene expressions of NLRP3, caspase-1 and IL-1β. In contrast, gene and protein expressions of IL-10 were reduced at 100 and 1000 μM H₂O₂. Protein expression of caspase-1 was higher in cultures of 100 μM H₂O₂ compared to controls. Treatment with Glybenclamide at 200 μM was used to prevent NLRP3 inflammasome activation. This concentration reduced protein expression of caspase-1 compared to culture with only H₂O₂ and control cultures.

Our results confirm that H₂O₂ induces oxidative stress on placental explants and demonstrate that cell responses to this stress involve inflammasome activation.