Electromagnetic field induced changes in lipid second messengers

Initial studies with a human hematopoietic cell line, TF-1, suggest multifarious effects of electromagnetic fields on lipid signal transduction. We have examined the effects of pulsed magnetic fields (2 T, 84 microseconds zero-to-peak haversine, 91 V/m induced electric field) on the cell cycle by flow cytometry. A 31% increase of cells in the G1 phase occurred concurrently with a 35% decrease of cells in S-phase, which suggests that doses of 30 or 40 pulses have an anti-proliferative effect. Changes in the lipid second messengers, diacylglycerol (DAG) and phosphatidic acid (PA) with stimuli of 2 T intensity were also dependent on the number of pulses. DAG production doubled with 30 pulses and tripled with 40 pulses, and PA levels were reduced to one third and one tenth of the original levels. Phospholipase D (PLD) up-regulation was assessed directly by the capacity of PLD to catalyze transphosphatidylation in the presence of alcohol. [3H]Phosphatidylethanol formed rapidly and continued to increase with concomitant decreases in [3H]PA and parallel generation of [3H]DAG. Propranolol, an inhibitor of PA phosphohydrolase, inhibited the formation of DAG in a dose-dependent manner with a marked increase in PA production. Examination of the kinetics of formation of [3H]choline and [3H]phosphocholine at different times after stimulation showed a rapid and consistent increase in [3H]choline, whereas [3H]phosphocholine increase was evident only 60 min after stimulation. Magnetic exposure also caused a shift in some molecular species patterns of DAG and PA which could be correlated with phosphatidylinositol, phosphatidylethanolamine and phosphatidylcholine molecular species decreases. Therefore, we propose that the PC-PLC pathway may be temporarily inactivated for a short period of time by exposure to pulsed stimuli, and the PC-PLD pathway is up-regulated based on: (1) cellular release of [3H]choline; (2) rapid intracellular formation of [3H]PA followed by [3H]DAG; (3)active transphosphatidylation; and (4) blockade of DAG formation by propranolol.