title: Impact of 17beta-estradiol on cytokine-mediated apoptotic effects in primary hippocampal and neocortical cell cultures.
abstract: Estrogens are developmental regulators of mitochondrial apoptotic pathway in the central nervous system, but little is known about their involvement in cytokine-induced apoptosis. In the present study, we evaluated effects of 17beta-estradiol on pro-inflammatory cytokine- and staurosporine-mediated activation of caspase-3 and LDH-release in primary neuronal/glial cell cultures of mouse hippocampal and neocortical cells at different stages of their development in vitro. Enzyme activities were determined with colorimetric methods 6 h, 14 h, 24 h, and 48 h after exposure to the apoptotic agents. Biochemical data were supported at the cellular level by Hoechst 33342 and MAP-2 stainings, which were carried out 48 h after the treatment. Cytokines (co-treatment with Il-1beta and TNFalpha; 1 ng/ml) increased caspase-3 activity in the hippocampal and neocortical cells up to over 200% of control values, and these effects were mostly observed on 2 and 7 days in vitro (DIV). Moderate, but significant cytokine-mediated increase in LDH-release was demonstrated in both tissues, especially on 7 and 12 DIV. Estradiol (100 nM) inhibited the activation of caspase-3 at early stage of development (2 DIV) in the hippocampal, but not in the neocortical cultures. The cytokine-induced activation of caspase-3 and LDH-release was inhibited by estradiol in estrogen receptor-independent way. These data point to a possible role of estrogens as non-estrogen receptor-related inhibitors of cytokine-activated apoptotic pathway in the developing central nervous system.
authors.affiliation: Department of Anatomy and Cell Biology, University of Ulm, Albert-Einstein-Alle 11, 89061 Ulm, Germany. kajta@if-pan.krakow.pl
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Kajta
authors.forename: Andreas
authors.initials: A
authors.lastname: Trotter
authors.forename: Wladyslaw
authors.initials: W
authors.lastname: Lasoń
authors.forename: Cordian
authors.initials: C
authors.lastname: Beyer
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles; D053148:Caspase 3 / Q000378:metabolism; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology / Q000502:physiology*; D003627:Data Interpretation, Statistical; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004958:Estradiol / Q000031:analogs & derivatives / Q000494:pharmacology*; D004965:Estrogen Antagonists / Q000494:pharmacology; D005260:Female; D005456:Fluorescent Dyes; D000077267:Fulvestrant; D006624:Hippocampus / Q000166:cytology / Q000502:physiology*; D053583:Interleukin-1beta / Q000494:pharmacology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008869:Microtubule-Associated Proteins / Q000187:drug effects / Q000378:metabolism; D019579:Neocortex / Q000166:cytology / Q000502:physiology*; D009457:Neuroglia / Q000187:drug effects / Q000201:enzymology; D009474:Neurons / Q000187:drug effects / Q000201:enzymology; D011247:Pregnancy; D019311:Staurosporine / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D001562:Benzimidazoles; D016207:Cytokines; D004791:Enzyme Inhibitors; D004965:Estrogen Antagonists; D005456:Fluorescent Dyes; D053583:Interleukin-1beta; D008869:Microtubule-Associated Proteins; D014409:Tumor Necrosis Factor-alpha; D000077267:Fulvestrant; D004958:Estradiol; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D019311:Staurosporine; C017807:bisbenzimide ethoxide trihydrochloride