| In Vitro | In vitro activity: Sodium salicylate acts as non-steroidal anti-inflammatory drug (NSAID), and induces apoptosis in cancer cells and also necrosis. Sodium salicylate inhibits the activation of NF-kappa B, and also inhibits NF-kappa B-dependent transcription from the Ig kappa enhancer and the human immunodeficiency virus (HIV) long terminal repeat (LTR) in transfected T cells. Sodium salicylate also induces apoptosis via p38 MAPK and inhibits TNF-induced JNK/stress-activated protein kinase activation.
Kinase Assay: Human purified COX-2 are and the cofactors Glutathione (5 mM),
Adrenaline (5 mM), and Hematin (1 μM) are dissolved in 50 mM Tris buffer
(pH 7.5). Hematin is first dissolved in a concentrated stock of 100 mM
in 1 M NaOH before being further diluted in Tris buffer. Enzyme
reactions are carried out in individual wells of 96-well plates with a
final reaction volume of 200 μL. Different concentrations of Sodium
Salicylate are added to the plate, followed by the addition of 10 units
of enzyme (180 μL). The plates are incubated at 37° for 30 min before
Arachidonic acid (10 nM to 30 μM) is added for a further 15 min. The
reaction is stopped by heating the plate to 100°C for 5 min. The 96-well
plate is then centrifuged at 10,000 × g for 10 min, and appropriated
samples are removed and added into the radioimmunoassay
Cell Assay: To assess the direct effect of Sodium Salicylate on COX-2 activity after induction has occurred, A549 cells are first treated with IL-1β for 24 hr, and the culture medium is replaced with DMEM containing different concentrations of Sodium Salicylate(10, 100 and 1000 μg/mL).
Cells are incubated at 37°C for 30 min. Arachidonic acid (1-30 μM) is
then added for 15 min, and the medium is removed for the measurement of
PGE2 |
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