Correlation between generation of nitric oxide and cell viability in human peripheral blood mononuclear cells and leukemic Jurkat T-cell line.
OBJECTIVE
To measure nitric oxide (NO) production in the form of nitrite derivative in relation to cell viability and apoptosis development in human peripheral blood mononuclear cells compared to that processes in human leukemic Jurkat T-cell line.


METHODS
Apoptosis was induced by dexamethasone (1 microg/ml) or NaNO(2) (7 microg/ml) added in the presence or absence of NO-synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) (27 microg/ml) during cell culturing. Cell viability was determined by trypan blue assay. Apoptosis was measured using DNA "ladder" assay.


RESULTS
Dexamethazone and NaNO(2) were shown to cause DNA "laddering" in both cell types. L-NAME prevented the appearance of apoptosis in both normal mononuclear cells of peripheral blood and leukemic Jurkat T-cell in the case of dexamethasone action, but it could not prevent it in the case of NaNO(2) action. The results of cell viability showed that both the dexamethasone and NaNO(2) significantly increased the percentage of dead cells. Their effect was better expressed in Jurkat T-cell line. The levels of nitrite production were higher in the leukemic T-cells comparing to such levels in the normal mononuclear cells.


CONCLUSIONS
Strong positive correlation was demonstrated between NO production and apoptosis development in both studied cell types, however leukemic Jurkat T-cell line responses were better expressed than such responses in normal mononuclear cells of peripheral blood. Potential significance of that correlation as well as possible mechanisms of appearing differences are discussed.
Levytskyy
Roman M
RM
Ivan Franko Lviv National University, Lviv, Ukraine.
Filyak
Yevgeniy Z
YZ
Stoika
Rostislav S
RS
D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival; D053938:DNA Fragmentation / Q000187:drug effects; D003907:Dexamethasone / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D019169:Jurkat Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D012977:Sodium Nitrite / Q000494:pharmacology
D004791:Enzyme Inhibitors; D009569:Nitric Oxide; D003907:Dexamethasone; D019001:Nitric Oxide Synthase; D012977:Sodium Nitrite; D019331:NG-Nitroarginine Methyl Ester