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

• Published in: Pregnancy hypertension Vol. 14; pp. 29 - 36
• Main Authors: Nunes, Priscila Rezeck, Peracoli, Maria Terezinha Serrao, Romao-Veiga, Mariana, Matias, Mariana Leticia, Ribeiro, Vanessa Rocha, Da Costa Fernandes, Celio Junior, Peracoli, Jose Carlos, Rodrigues, Jose Ricardo, De Oliveira, Leandro
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-10-2018
• Subjects: Caspase 1 - drug effects; Caspase 1 - metabolism; Cytokines; Female; Humans; Hydrogen Peroxide - pharmacology; Inflammasome; Inflammasomes - drug effects; Inflammasomes - metabolism; Oxidative stress; Oxidative Stress - drug effects; Placenta - drug effects; Placenta - metabolism; Placental explants; Polymerase Chain Reaction; Pre-Eclampsia - metabolism; Pregnancy; Tissue Culture Techniques; Oxidative stress; Inflammasome; Cytokines; Placental explants
• ISSN: 2210-7789; 2210-7797
• DOI: 10.1016/j.preghy.2018.07.006

•Oxidative stress activate inflammasome, with pro-inflammatory cytokine production.•Inflammasome activation could be a response to oxidative stress.•Antioxidants try to combat oxidative stress generated by H2O2 in 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.