Cell experiment:

The myocytes are plated on 35 mm plastic culture dishes at a density of approximately 9×105 cells/mL (1.8×106 cells/dish). The cells are kept in culture for 6 d at 37℃ and 5% CO2 under aerobic conditions in a carbon dioxide incubator. During this period the cells are beating spontaneously at an approximate frequency of 20 to 40 beats/min. Each culture dish contains 0.9+0.2 mg protein. The antioxidant ICI 211965 is added to the culture medium at a final concentration of 0.1% ethanol. Cell damage is determined by leakage of lactate dehydrogenase (LD) to the medium, and is analyzed after 0.5, 1.0, 2.0, 3.0, and 5.0 h[1].

ICI 211965 is a selective and orally potent 5-Lipoxygenase (5-LPO) inhibitor.

ICI 211965 is a selective and orally potent 5-Lipoxygenase (5-LPO) inhibitor. ICI 211965 reduces the leakage of lactate dehydrogenase (LD) and lipid peroxidation at concentrations related to its antioxidative effect and not to its 5-1ipoxygenase-inhibiting effect[1].

[1]. Crawley GC, et al. Methoxytetrahydropyrans. A new series of selective and orally potent 5-lipoxygenase inhibitors. J Med Chem. 1992 Jul 10;35(14):2600-9.