Tert -Butyl Hydroperoxide Stimulated Apoptosis Independent of Prostaglandin E 2 and IL-6 in the HTR-8/SVneo Human Placen
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ORIGINAL ARTICLE
Tert-Butyl Hydroperoxide Stimulated Apoptosis Independent of Prostaglandin E2 and IL-6 in the HTR-8/SVneo Human Placental Cell Line Rita Loch-Caruso 1
&
Cassandra S. Korte 1,2
&
Kelly A. Hogan 1,3
&
Sarah Liao 1,4 & Craig Harris 1
Received: 3 December 2019 / Revised: 10 May 2020 / Accepted: 3 June 2020 # Society for Reproductive Investigation 2020
Abstract Significant gaps exist in our knowledge of how cellular redox status, sometimes referred to as oxidative stress, impacts placental trophoblasts. The present study used tert-butyl hydroperoxide (TBHP) as a known generator of reactive oxygen species (ROS) in the extravillous trophoblast cell line HTR-8/SVneo to examine the role of cellular redox disruption of prostaglandin E2 (PGE2) and the cytokine IL-6 in cell death. Cells were exposed to 0, 12.5, 25, or 50 μM TBHP for 4, 8, and 24 h to ascertain effects on cell viability, caspase 3/7 activity, PGE2 release, PTGS2 mRNA expression, and IL-6 release. Experiments with inhibitors included the cyclooxygenase inhibitor indomethacin, mitogen-activated protein kinase inhibitors (PD169316, U0126, or SP600125), or treatments to counter expected consequences of TBHP-stimulated generation of ROS (deferoxamine [DFO], butylated hydroxyanisole [BHA], and N,N′-diphenyl-1,4-phenylenediamine [DPPD]) using 24-h exposure to 50 μM TBHP. Cell viability, measured by ATP content, decreased 24% relative to controls with a 24-h exposure to 50 μM TBHP, but not at lower TBHP concentrations nor at earlier time points. Exposure to 50 μM TBHP increased caspase 3/7 activity, an indicator of apoptosis, after 8 and 24 h. Antioxidant treatment markedly reduced TBHP-stimulated caspase 3/7 activity, PGE2 release, and IL-6 release. TBHP-stimulated IL-6 release was blocked by PD169316 but unaltered by indomethacin. These data suggest that TBHPstimulated IL-6 release and caspase 3/7 activation were independent of PGE2 yet were interrupted by treatments with known antioxidant properties, providing new insight into relationships between PGE2, IL-6, and apoptosis under conditions of chemically induced cellular oxidation. Keywords Oxidative stress . Reactive oxygen species . Trophoblasts . Apoptosis . Prostaglandins
Introduction Nutrient and oxygen exchange between mother and fetus relies on the establishment of the placental maternal-fetal interface. The extravillous trophoblasts are critical for placentation
success with their proliferation, migration, and invasion of maternal uterine tissues. These trophoblast functions are regulated by factors such as hormones, prostaglandins, and cytokines [1, 2]. Disrupted regulation of extravillous trophoblast functions during placentation can lead to placental
Rita Loch-Caruso and Cassandra S. Korte contributed equally to this work. * Rita Loch-Caruso [email protected]
Craig Harris [email protected]
Cassandra S. Korte [email protected]
1
Department of Environmental Health Sciences, School of Public Health, University of Michigan, Ann Arbor, MI 48109-2029, USA
Kelly A. Hogan hogan.kelly@m
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