Induction of three-dimensional assembly and increase in apoptosis of human endothelial cells by simulated microgravity: Impact of vascular endothelial growth factor

被引:122
作者
Infanger, M.
Kossmehl, P.
Shakibaei, M.
Baatout, S.
Witzing, A.
Grosse, J.
Bauer, J.
Cogoli, A.
Faramarzi, S.
Derradji, H.
Neefs, M.
Paul, M.
Grimm, D.
机构
[1] Charit Univ Med, Inst Clin Pharmacol & Toxicol, Sch Med, Ctr Space Med, D-14195 Berlin, Germany
[2] Charite, Sch Med, Dept Trauma & Reconstruct Surg, Benjamin Franklin Med Ctr,Ctr Space Med, D-12200 Berlin, Germany
[3] Univ Munich, Inst Anat, D-80336 Munich, Germany
[4] Charite, Sch Med, Inst Anat, D-14195 Berlin, Germany
[5] CEN SCK, Belgian Nucl Res Ctr, Radiobiol Lab, B-2400 Mol, Belgium
[6] Max Planck Inst Biochem, D-82152 Martinsried, Germany
[7] ETH, Zero G Lifetec GmbH, CH-8005 Zurich, Switzerland
关键词
apoptosis; endothelial cells; extracellular matrix proteins; microgravity; VEGF; weightlessness;
D O I
10.1007/s10495-006-5697-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Endothelial cells play a crucial role in the pathogenesis of many diseases and are highly sensitive to low gravity conditions. Using a three-dimensional random positioning machine (clinostat) we investigated effects of simulated weightlessness on the human EA.hy926 cell line (4, 12, 24, 48 and 72 h) and addressed the impact of exposure to VEGF (10 ng/ml). Simulated microgravity resulted in an increase in extracellular matrix proteins (ECMP) and altered cytoskeletal components such as microtubules (alpha-tubulin) and intermediate filaments (cytokeratin). Within the initial 4 h, both simulated microgravity and VEGF, alone, enhanced the expression of ECMP (collagen type I, fibronectin, osteopontin, laminin) and flk-1 protein. Synergistic effects between microgravity and VEGF were not seen. After 12 h, microgravity further enhanced all proteins mentioned above. Moreover, clinorotated endothelial cells showed morphological and biochemical signs of apoptosis after 4 h, which were further increased after 72 h. VEGF significantly attenuated apoptosis as demonstrated by DAPI staining, TUNEL flow cytometry and electron microscopy. Caspase-3, Bax, Fas, and 85-kDa apoptosis-related cleavage fragments were clearly reduced by VEGF. After 72 h, most surviving endothelial cells had assembled to three-dimensional tubular structures. Simulated weightlessness induced apoptosis and increased the amount of ECMP. VEGF develops a cell-protective influence on endothelial cells exposed to simulated microgravity.
引用
收藏
页码:749 / 764
页数:16
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