Flow cytometric estimation of the plasma membrane diversity of bone marrow cells in mice treated with WR-2721 and cyclophosphamide.
The effects of S-2-/3-aminopropylamino/ethyl phosphorothioic acid (WR-2721, Amifostine) and cyclophosphamide (CP) on the cell surface exposure of phosphatidylserine (PS) and the plasma membrane impairment of bone marrow cells were assessed by flow cytometry assay with fluoresceinated annexin V (annexin V - FITC) and propidium iodide (PI). During the 96 h-period after treatment of adult male Swiss mice with WR-2721 (400 mg/kg b.wt.) and CP (200 mg/kg b.wt.), bone marrow cells expressing PS on the outer leaflet of the plasma membrane, which bound annexin V, and cells with a compromised cell membrane, which allowed PI to bind to the cellular DNA, were analysed. Temporary changes in the frequency of early apoptotic cells (annexin V - FITC positive/PI negative), late apoptotic and necrotic cells (annexin V - FITC positive/PI positive), and the number of live cells (annexin V - FITC negative/PI negative), were dependent on the drug(s) given. Application of CP distinctly triggered apoptotic and necrotic cell death, and WR-2721 pre-treatment of mice affected cell death induced by CP, causing reduction of the number of apoptotic and necrotic cells. The chemoprotective action of WR-2721 against PS externalisation and the plasma membrane impairment of normal bone marrow cells was shown.
Mazur
Lidia
L
Laboratory of Experimental Hematology and Toxicology, Department of Animal Physiology, Institute of Zoology, Jagiellonian University, R. Ingardena 6, 30-060, Cracow, Poland. maz@zuk.iz.uj.edu.pl
Augustynek
Anna
A
Bochenek
Michał
M
D004999:Amifostine / Q000494:pharmacology*; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000473:pathology; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D003520:Cyclophosphamide / Q000494:pharmacology*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D008297:Male; D051379:Mice; D010718:Phosphatidylserines / Q000378:metabolism; D011419:Propidium; D013997:Time Factors
D017304:Annexin A5; D010718:Phosphatidylserines; D011419:Propidium; D003520:Cyclophosphamide; D016650:Fluorescein-5-isothiocyanate; D004999:Amifostine