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http://dx.doi.org/10.5352/JLS.2009.19.5.598

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.  

Kwon, Gun-Rok (Dept. of Oriental Medicine)
Moon, Il-Soo (Department of Anatomy, Dongguk University)
Lee, Won-Chul (Pusan National University School of Oriental Medicine)
Publication Information
Journal of Life Science / v.19, no.5, 2009 , pp. 598-606 More about this Journal
Abstract
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${\sim}$5, 5.0 ${\mu}g/ml$) on day in vitro 12(DIV12), given a hypoxic shock (2% $O_2$/5% $CO_2$, $37^{\circ}C$, 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${\sim}$15% increase in viability; p<0.01) at 1.0 and 2.5 ${\mu}g/ml$ in normoxia and hypoxia. Measurement of ROS production by $H_2DCF-DA$ 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.
Keywords
Cortical cell culture; hypoxia; MMP; Pinelliae Rhizoma; ROS;
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