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KMID : 0613820090190050598
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Journal of Life Science
2009 Volume.19 No. 5 p.598 ~ p.606
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Neuroprotective Effects of Pinelliae Rhizoma Water-Extract by Suppression of Reactive Oxygen Species and Mitochondrial Membrane Potential Loss in a Hypoxic Model of Cultured Rat Cortical Cells
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Kwon Gun-Rok
Moon Il-Soo
Lee Won-Chul
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Abstract
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Oxidative stress by free radicals is a major cause of neuronal cell death. Excitotoxicity in hypoxia/ischemia causes an increase in reactive oxygen species (ROS) and a loss of mitochondrial membrane potential (MMP), resulting in dysfunction of the mitochondria and cell death. Pinelliae Rhizoma (PR) is a traditional medicine for incipient stroke. We investigated the effects of PR Water-Extract on the modulation of ROS and MMP in a hypoxic model using cultured rat cortical cells. PR Water-Extract was added to the culture medium at various concentrations (0.25～5, 5.0 ㎍/ml) on day in vitro 12(DIV12), given a hypoxic shock (2% O₂/5% CO₂, 37℃, 3 hr), and cell viability was assessed on DIV15 by Lactate Dehydrogenase Assay (LDH assays). PR Water-Extract showed a statistically significant effect on neuroprotection (10～15% increase in viability; p＜0.01) at 1.0 and 2.5 μg/ml in normoxia and hypoxia. Measurement of ROS production by H2DCF-DA stainings showed that PR Water-Extract efficiently reduced the number and intensity of ROS-producing neurons, especially at 1 hr post shock and DIV15. When MMP was measured by JC-1 stainings, PR Water-Extract efficiently maintained high-energy charged mitochondria. These results indicate that PR Water-Extract protects neurons in hypoxia by preventing ROS production and preserving the cellular energy level.
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KEYWORD
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Cortical cell culture, hypoxia, MMP, Pinelliae Rhizoma, ROS
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