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Full Text Pdf Eastern Biologist No. 4 2015 Protective Effects of Conjugated Equine Estrogens and 17-β Estradiol on Oxidatively Stressed Astrocytes Whitley E. Grimes and Kathleen S. Hughes The Eastern Biologist . ♦ A peer-reviewed journal that publishes original articles focused on the many diverse disciplines of biological research, except for the natural history science disciplines (ISSN 2165-6657 [online]). ♦ Subject areas - The journal welcomes manuscripts based on original research and observations, as well as research summaries and general interest articles. Subject areas include, but are not limited to, biochemistry, biotechnology, cell biology, developmental biology, genetics and genomics, immunology, microbiology, molecular evolution, neurobiology, parasitology, physiology, toxicology, as well as scientific pedagogy. ♦ Offers article-by-article online publication for prompt distribution to a global audience ♦ Offers authors the option of publishing large files such as data tables, audio and video clips, and even PowerPoint presentations as online supplemental files. ♦ Special issues - The Eastern Biologist welcomes proposals for special issues that are based on conference proceedings or on a series of invitational articles. Special issue editors can rely on the publisher’s years of experiences in efficiently handling most details relating to the publication of special issues. ♦ Indexing - As is the case with Eagle Hill's other journals, the Eastern Biologist is expected to be fully indexed in Elsevier, Thomson Reuters, Proquest, EBSCO, Google Scholar, and other databases. ♦ The journal staff is pleased to discuss ideas for manuscripts and to assist during all stages of manuscript preparation. The journal has a mandatory page charge to help defray a portion of the costs of publishing the manuscript. Instructions for Authors are available online on the journal’s website (http://www.eaglehill. us/ebio) or by e-mail ([email protected]). ♦ Co-published with the Northeastern Naturalist, Southeastern Naturalist, Caribbean Naturalist, and Urban Naturalist. Each journal has a separate Board of Editors. ♦ Available online in full-text version on the journal's website (http://www. eaglehill.us/ebio). Arrangements for inclusion in other BioOne databases are being pursued. Cover Photograph: Sample media and disposable vessels used in cultured astrocyte studies EASTERN BIOLOGIST Board of Editors Brian Burnes, Department of Biology, University of West Alabama, Livingston, AL Judy Awong-Taylor, School of Science and Technology, Georgia Gwinnett College, Lawrenceville, GA Chris R. Gissendanner, College of Pharmacy, University of Louisiana, Monroe, LA Robert Gregerson, College of Science and Technology, Armstrong Atlantic State Univ., Savannah, GA Jill Lee, Eagle Hill Institute, Steuben, ME ... Assistant Editor Tim H. Lindblom, Math and Science Division, Lyon College, Batesville, AR ... Managing Editor Joerg-Henner Lotze, Eagle Hill Institute, Steuben, ME ... Publisher Christi Magrath, Department of Biological and Environmental Sciences, Troy University, Troy, AL Marilyn S. Mayer, Eagle Hill Institute, Steuben, ME ... Production Editor Scott Merrell, Department of Microbiology and Immunology, Uniformed Services, University of the Health Sciences, Bethesda, MD Ashley B. Morris, Department of Biology, Middle Tennessee State University, Murfreesboro, TN J. Kenneth Shull, Department of Biology, Appalachian State University, Boone, NC (Emeritus) Stephanie R. Songer, Department of Biology, North Georgia College and State University, Dahlonega, GA Darla J. Wise, Biology Department, Concord University, Athens, WV Dwayne A. Wise, Department of Biological Sciences, Mississippi State University, Mississippi State, MS The Eastern Biologist (ISSN # 2165-6657) is published by the Eagle Hill Institute, PO Box 9, 59 Eagle Hill Road, Steuben, ME 04680-0009. Phone 207-546-2821, FAX 207-546-3042. E-mail: [email protected]. Webpage: http://www.eaglehill.us/ebio. Copyright © 2014, all rights reserved. Published on an article by article basis. Special issue proposals are welcome. Eastern Biologist is an open access journal. Authors: submission guidelines are available at http://www.eaglehill.us/ebio. Co-published jour- nals: The Northeastern Naturalist, Southeastern Naturalist, Caribbean Naturalist, and Urban Naturalist, each with a separate Board of Editors. The Eagle Hill Institute is a tax exempt 501(c)(3) nonprofit corporation of the State of Maine (Federal ID # 010379899). 2015 Eastern Biologist No. 4 2015 4:1–10 W.E.EASTERN Grimes and BIOLOGIST K.S. Hughes Protective Effects of Conjugated Equine Estrogens and 17-β Estradiol on Oxidatively Stressed Astrocytes Whitley E. Grimes1 and Kathleen S. Hughes1,* Abstract - Conjugated equine estrogens (CEEs), estradiol, and progestins are used in com- mon hormone replacement prescriptions for menopausal and post-menopausal women. While studies have linked 17-β estradiol to cell viability, little is known about the effects of individual CEEs (equilin and equilenin) during oxidative stress. This study sought to examine the protective effects of pretreating astrocytes with varying concentrations of equilin, equilenin, or 17-β estradiol, on the impact of oxidative stress on cell viability. We hypothesized that astrocytes pretreated with either CEE would not be protected from oxida- tive stress induced by exposure to hydrogen peroxide (H2O2), whereas cells pretreated with 17-β estradiol would be. MTT assay results revealed that the viability of untreated cells exposed to 600 μM H2O2 for either 1 h or 24 h was significantly reduced compared to the viability of unstressed cells. The viability of astrocytes pretreated with 10 nM – 10 μM of either CEE (equilin or equilenin) for 30 min prior to being stressed by a 1-h exposure to H2O2 did not differ from the viability of unstressed cells, indicating that CEE pretreatment provided some level of protection from oxidative stress. However, neither the CEEs- nor 17-β estradiol-pretreatment consistently protected astrocytes exposed to H2O2 for 24 h. Fu- ture research should examine the effect of increased duration of pretreatments, the effects of pretreatments on neuronal-like cell lines, and other measures of oxidative stress. Introduction Hormone replacement therapy (HRT) is an approved method of treatment for menopausal and post-menopausal women, and clinical trials reveal both detrimen- tal and beneficial effects of HRT (Howard, B.V. and J.E. Rossouw. 2013, Prentice, R.L. 2014). While hormone replacement therapy can decrease some of the negative side effects of menopause, long-term HRT is associated with dementia, Alzheimer’s disease, and an increased risk of uterine cancer (Rousseau 2010). Estrogens are pri- marily female sex hormones that bind to estrogen receptors (ER) in the cell. Much of the research on estrogen signaling focuses on its traditional pathway of binding to a receptor such as the estrogen receptor alpha in the cytoplasm, translocating to the nucleus, and affecting transcription (McDevitt, M.A et. al 2008). Estrogens are also able to elicit faster effects by binding to transmembrane G protein-coupled receptors and activating signaling pathways (Weatherman 2006). Previous research identified that estrogen pretreatment for only 30 min protected cells from oxidative stress, which points to non-nuclear signaling (Yu et al. 2004). There are three estrogen classes that naturally occur in women: estrone, estradiol, and estriol. Estradiol levels in the body drop from 15–350 pg/mL in premenopausal Department of Biology, Columbus State University, Columbus, GA 31907. *Corresponding author – [email protected], 706-568-2325 Manuscript Editor: Christopher Gissendanner 1 2015 Eastern Biologist No. 4 W.E. Grimes and K.S. Hughes women to less than 10 pg/mL in postmenopausal women. Estrone levels in the body drop from 17–200 pg/mL for premenopausal women to 7–40 pg/mL in postmenopaus- al women. Estriol levels in premenopausal women are 8–11 pg/ml (Rotti et al. 1975). Estradiol, specifically 17-β estradiol, plays a key role in the protection of neurological faculties and cardiac functions (Ito et al. 2006, Zhao et al. 2006). Seventeen-β estradiol is thought to contribute to cell protection through tran- scriptional regulation, though evidence points to action through transmembrane G protein-coupled receptors as well. Evidence shows that 17-β estradiol is also known to protect against oxidative stress, especially oxidative stress-induced neuronal cell apoptosis in relation to Alzheimer’s disease (Behl et al. 1997). Prescribed medications for hormone replacement therapy can include multiple types of estrogens such as estradiol, estrone, the conjugated equine estrogens (CEEs) equilin, and equilenin, as well as progestins. Despite the seemingly positive benefits of endogenous 17-β estradiol, the effect can be deleterious when combined with CEEs. Conjugated equine estrogens are taken from the urine of pregnant mares and are not found in humans. Two types available for study are equilin and equilenin. CEEs have been shown to minimize the side-effects of menopause such as hot flashes, night sweats, and mood swings (Brunner, R.L. et al. 2010). Unlike 17-β estradiol, they do not protect against neuronal intracellular ATP loss when de- livered at 300–1000 pg/ml in vitro (Zhao et al. 2006). However, when several CEEs are administered together, the neuroprotective benefits such as protection from stroke and a lessened chance for Alzheimer’s are measurable (Zhao et al. 2006).
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