Electrochemically Reduced Water Protects Neural Cells from Oxidative Damage

Figure 7

Suppressive effect of ERW on glutamate-induced Ca²⁺ influx in MCCNP cells. Intracellular Ca²⁺ was measured with Fluo-3 AM, a membrane-permeable calcium sensitive dye. Internalized Fluo-3 AM is hydrolyzed by intracellular esterases liberating Flou-3 which can react with intracellular free Ca²⁺ ions forming a fluorescent complex. After pretreatment with glutamate (0, 0.1 or 1.0 mM) in the medium made with ERW or MQ-water for 15 min, MCCNP cells were stained with Fluo-3 AM for 20 min and post-incubated with FBS/HBSS medium. A fluorescence microscope was used to detect intracellular fluorescence and the images were recorded by a digital camera. Digital images were converted to numerical data with the aid of the NIH image analysis program and then analyzed by Excel software. Representative histograms of triplicate independent experiments are shown.