Document Server@UHasselt >
Research >
Research publications >

Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/18639

Title: Dobutamine mediates cytoprotection by induction of heat shock protein 70 in vitro
Authors: Roesslein, M.
Froehlich, C.
JANS, Frank
Piegeler, T.
Goebel, U.
Loop, T.
Issue Date: 2014
Citation: LIFE SCIENCES, 98 (2), p. 88-95
Abstract: Aims: Dobutamine is cytoprotective when applied before a subsequent stress. However, the underlying molecular mechanism is unknown. Dobutamine also inhibits nuclear factor (NF)-κB in human T lymphocytes. Other inhibitors of NF-κB induce a so-called heat shock response. We hypothesized that dobutamine mediates protection from apoptotic cell death by the induction of a heat shock response. Main methods: Jurkat T lymphoma cells were preincubated with dobutamine (0.1, 0.5 mM) before the induction of apoptosis (staurosporine, 2 μM). DNA-binding of heat shock factor (HSF)-1 was analyzed by electrophoretic mobility shift assay, mRNA-expression of heat shock protein (hsp)70 and hsp90 by Northern Blot, activity of caspase-3 by fluorogenic caspase activity assay and cleavage of pro-caspase-3 by Western Blot. Apoptosis was assessed by flow cytometry after annexin V-fluorescein isothiocyanate staining. Hsp70 and hsp90 were inhibited using N-formyl-3,4-methylenedioxy-benzylidene-gamma-butyrolaetam and 17-allylamino-17-demethoxygeldana-mycin, respectively. All data are given as median and 25/75% percentile. Key findings: Pre-incubation with dobutamine inhibited staurosporine-induced annexin V-fluorescence (28 [20–32] % vs. 12 [9–15] % for dobutamine 0.1 mM and 7 [5–12] % for dobutamine 0.5 mM, p b 0.001), cleavage of procaspase-3 as well as caspase-3-like activity (0.46 [0.40–0.48] vs. 0.32 [0.27–0.39] for Dobutamine 0.1 mM and 0.20 [0.19–0.23] for Dobutamine 0.5 mM, p b 0.01). Dobutamine induced DNA-binding of HSF-1 and mRNAexpression of hsp70 and hsp90. While inhibition of Hsp90 had no effect, inhibition of Hsp70 increased the number of annexin V-positive cells (33 [32–36] % vs. 18 [16–24] %) and caspase-3-like activity (0.21 [0.19–0.23] vs. 0.16 [0.13–0.17], p b 0.05). Significance: Dobutamine protects from apoptotic cell death via the induction of Hsp70.
Notes: Corresponding author at: University Medical Center Department of Anaesthesiology and Critical Care Medicine Hugstetter Str. 55 D-79106 Freiburg, Germany. Tel./fax: +49 761 270 23 380/930. E-mail address: martin.roesslein@uniklinik-freiburg.de (M. Roesslein).
URI: http://hdl.handle.net/1942/18639
DOI: 10.1016/j.lfs.2014.01.005
ISI #: 000332443000005
ISSN: 0024-3205
Category: A1
Type: Journal Contribution
Validation: ecoom, 2015
Appears in Collections: Research publications

Files in This Item:

Description SizeFormat
N/A746.24 kBAdobe PDF

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.