ROS-challenged keratinocytes as a new model for oxidative stress-mediated skin diseases

Journal of Cellular Biochemistry
Victoria BaryginaClaudia Fiorillo

Abstract

In the current study, the effects of the reactive oxygen species (ROS) generator 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) on extracellular and intracellular ROS production in human keratinocytes (HACAT) were studied. AAPH is a water-soluble compound able to generate ROS at known and constant rates at 37°C. The short treatment (2 h) with AAPH brought a significant dose-dependent increase in NADPH oxidase activity in intact keratinocytes. The long-term treatment (24 h) with AAPH led to a persistent increase in NADPH oxidase activity for up to 48 hour following the AAPH removal from cell incubation medium. ROS and nitric oxide levels, lipoperoxidation, intracellular calcium, mitochondrial superoxide production, and membrane potential were significantly modified in AAPH-treated HACAT. Superoxide dismutase (SOD) and/or catalase addition to HACAT revealed that untreated keratinocytes produce mostly superoxide anion (O 2- ), while AAPH-treated keratinocytes overproduce hydrogen peroxide (H 2 O 2 ) in extracellular medium. H 2 O 2 is particularly stable and plays important roles in several cell signaling pathways. Taken together, our findings suggest a cost-effective and easily reproducible in vitro model of stressed human ...Continue Reading

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Citations

Jun 2, 2020·Frontiers in Medicine·Giovanna G AltobelliVincenzo Cimini
Aug 11, 2020·Free Radical Biology & Medicine·Camilo López-AlarcónMichael J Davies
Sep 26, 2021·Clinical and Experimental Immunology·Alessandra BettiolClaudia Fiorillo

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