• The Korean Journal of Physiology and Pharmacology
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• v.3 no.3
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• pp.357-364
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• 1999

We investigated the role of $Ca^{2+}$ and protein kinases/phosphatases in the stimulatory effect of insulin on glucose transport. In isolated rat adipocytes, the simple omission of $CaCl_2$ from the incubation medium significantly reduced, but did not abolish, insulin-stimulated 2-deoxy glucose (2-DG) uptake. Pre-loading adipocytes with intracellular $Ca^{2+}$ chelator, 5,5'-dimethyl bis (o-aminophenoxy)ethane-N,N,N'N' tetraacetic acetoxymethyl ester (5,5'-dimethyl BAPTA/AM) completely blocked the stimulation. Insulin raised intracellular $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ about 1.7 times the basal level of $72{\pm}5$ nM, and 5,5'-dimethyl BAPTA/AM kept it constant at the basal level. This correlation between insulin-induced increases in 2-DG uptake and $[Ca^{2+}]_i$ indicates that the elevation of $[Ca^{2+}]_i$ may be prerequisite for the stimulation of glucose transport. Studies with inhibitors (ML-9, KN-62, cyclosporin A) of $Ca^{2+}-calmodulin$ dependent protein kinases/phosphatases also indicate an involvement of intracellular $Ca^{2+}.$ Additional studies with okadaic acid and calyculin A, protein phosphatase-1 (PP-1) and 2A (PP-2A) inhibitors, indicate an involvement of PP-1 in insulin action on 2-DG uptake. These results indicate an involvement of $Ca^{2+}-dependent$ signaling pathway in insulin action on glucose transport.

• BMB Reports
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• v.37 no.2
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• pp.239-245
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• 2004

We have previously demonstrated that the redox reactant pyruvate prevents apoptosis in the oxidant model of bovine pulmonary artery endothelial cells (BPAEC), and that the anti-apoptotic mechanism of pyruvate is mediated in part via the mitochondrial matrix compartment. However, cytosolic mechanisms for the cytoprotective feature of pyruvate remain to be elucidated. This study investigated the pyruvate protection against endothelial cytotoxicity when the glycolysis inhibitor 2-deoxy-D-glucose (2DG) was applied to BPAEC. Millimolar 2DG blocked the cellular glucose uptake in a concentration- and time-dependent manner with >85% inhibition at $\geq$5 mM within 24 h. The addition of 2DG evoked BPAEC cytotoxicity with a substantial increase in lipid peroxidation and a marked decrease in intracellular total glutathione. Exogenous pyruvate partially prevented the 2DG-induced cell damage with increasing viability of BPAEC by 25-30%, and the total glutathione was also modestly increased. In contrast, 10 mM L-lactate, as a cytosolic reductant, had no effect on the cytotoxicity and lipid peroxidation that are evoked by 2DG. These results suggest that 2DG toxicity may be a consequence of the diminished potential of glutathione antioxidant, which was partially restored by exogenous pyruvate but not L-lactate. Therefore, pyruvate qualifies as a cytoprotective agent for strategies that attenuate the metabolic dysfunction of the endothelium, and cellular glucose oxidation is required for the functioning of the cytosolic glutathione/NADPH redox system.

• Clinical and Experimental Pediatrics
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• v.51 no.10
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• pp.1112-1117
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• 2008

Purpose : We intended to observe cell death and apoptotic changes in neurons in organotypic hippocampal slice cultures following oxygen-glucose deprivation (OGD), using propidium iodide (PI) uptake, Fluoro-Jade (FJ) staining, TUNEL staining and immunofluorescent staining for caspase-3. Methods : The hippocampus of 7-day-old rats was cut into $350{\mu}m$ slices. The slices were cultured for 10 d (date in vitro, DIV 10) and and exposed to OGD for 60 min at DIV 10. They were then incubated for reperfusion under normoxic conditions for an additional 48 h. Fluorescence of PI uptake was observed at predetermined intervals, and the cell death percentage was recorded. At 24 h following OGD, the slices were Cryo-cut into $15{\mu}m$ thicknesses, and Fluoro-Jade staining, TUNEL staining, and immunofluorescence staining for caspase-3 were performed. Results : 1) PI uptake was restricted to the pyramidal cell layer and DG in the slices after OGD. The fluorescent intensities of PI increased from 6 to 48 h during the reperfusion stage. The cell death percentage significantly increased time-dependently in CA1 and DG following OGD (P<0.05). 2) At 24 h after OGD, many FJ positive cells were detected in CA1 and DG. Some neurons had distinct nuclei and processes while others had fragmented nuclei and disrupted processes in CA1. TUNEL and immunofluorescent staining for caspase-3 showed increased expression of TUNEL labeling and caspase-3 in CA1 and DG at 24 h after OGD. Conclusion : The numerous dead cells in the slice cultures after OGD tended to display apoptotic changes mediated by the activation of caspase-3.

• Proceedings of the Membrane Society of Korea Conference
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• 1999.07a
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• pp.115-115
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• 1999

In this work a new process was developed for the sulfonation of the chemicallly stable engineering polymer polyethersulfone as membrane materials for electrodialysis or a flow battery applications. Commercially available polyethersulfone polymer was partially sulfonated using a CSA sulfonating agent in a dichloromethane solvent, which sulfonated polyethersulfone with various sulfonation levels have been prepared. Sulfonated polyethersulfone (SPES) membranes with different ion capacities were prepared for the purpose of identifying cation exchange membrane properties, in an attempt to find a low cost replacement for Nafion, which most of the perfluorinated membranes, known to exhibit a prolonged service life, are expensive and difficult to process. The following features were determined: the degree of sulfonation, water uptake, thermal analysis, and electrochemical properties such as ion exchange capacities, resistivity, selectivity of ion permeation. The surface of the cation exchange membranes, decomposed with the H202-treatment, were observed by using scanning electron microscope. The area resistivities of SPES mebranes in 5N-NaOH decreased from $2,150{\;}{\Omega}-cm2$ to less than $15{\Omega}-cm2$ as the ion exchange capacity (IEC) increased from 0.62 to 1.73 millieequivlants per dry gram(meq/dg).eq/dg).

• Clinical and Experimental Pediatrics
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• v.52 no.5
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• pp.588-593
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• 2009

Purpose : To investigate whether growth hormone (GH) has a protective effect on neurons in hippocampal slice cultures of neonatal rats exposed to oxygen-glucose deprivation (OGD). Methods : Cultured hippocampal slices of 7-day-old rats were exposed to OGD for 60 min. Then, the slices were immediately treated with three doses of GH (5, 50, or $500{\mu}M$) in media. The relative fluorescent densities of propidium iodide (PI) uptake in the slices and relative lactate dehydrogenase (LDH) activities in the media were determined and compared between each GH- treated group of slices and untreated slices (control) at 12 and 24 h after OGD. Immunofluorescent staining for caspase-3 and TUNEL staining were performed to observe the effect of GH on apoptotic neuronal death. Results : The relative fluorescent densities of PI uptake in CA1 and dentate gyrus (DG) of the hippocampal slices in each GH-treated group were not significantly different from those in the untreated slices at 12 and 24 h after OGD (P>0.05). Treatment with GH could reduce the relative LDH activities in the media of the GH-treated groups only at 12 h after OGD (P<0.05). Expression of caspase-3 and TUNEL positivity in CA1 and DG of the slices treated with 50-iM GH were not different from those of the untreated slices at 12 and 24 h after OGD. Conclusion : Treatment of hippocampal slice cultures with GH after OGD does not show a definitive protective effect on neuronal death but can reduce the LDH efflux of the slices in media at 12 h after OGD.

• Journal of the Korean Institute of Landscape Architecture
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• v.42 no.5
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• pp.13-21
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• 2014

This study generated regression models to quantify storage and annual uptake of carbon from five native landscape tree species through a direct harvesting method, and established essential information to estimate carbon reduction effects from urban greenspaces. Tree species for the study included the Chionanthus retusus, Prunus armeniaca, Abies holophylla, Cornus officinalis, and Taxus cuspidata, which are usually planted in cities of middle Korea, but for which no information on carbon reduction is available. Ten tree individuals for each species were sampled reflecting various stem diameter sizes at a given interval. The study measured biomass for each part including the roots of sample trees to compute total carbon storage per tree. The annual carbon uptake per tree was quantified by analyzing the radial growth rates of stem samples at breast height or ground level. Regression models were developed using diameter at breast height (dbh) or ground level (dg) as an independent variable to easily estimate storage and annual uptake of carbon per tree for each species. All the regression models showed high fitness with $r^2$ values of 0.92~0.99. Storage and annual uptake of carbon from a tree with dbh of 10 cm were greatest with C. retusus (20.0 kg and 5.9 kg/yr, respectively), followed by P. armeniaca (17.5 kg and 4.5 kg/yr) and A. holophylla (13.2kg and 1.8 kg/yr) in order. A C. officinalis tree and T. cuspidata tree with dg of 10 cm stored 9.3 and 6.3 kg of carbon and annually sequestered 3.2 and 0.6 kg, respectively. The above-mentioned carbon storage equaled the amount of carbon emitted from gasoline consumption of about 23~35 L for C. retusus, P. armeniaca, and A. holophylla, and 11~16 L for C. officinalis and T. cuspidata. A tree with the diameter size of 10 cm annually offset carbon emissions from gasoline use of about 6~10 L for C. retusus, P. armeniaca, and C. officinalis, and 1~3 L for A. holophylla and T. cuspidata. The study breaks new ground to easily quantify biomass and carbon reduction for the tree species by overcoming difficulties in direct cutting and root digging of urban landscape trees.

• Journal of the Korean Institute of Landscape Architecture
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• v.47 no.3
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• pp.31-38
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• 2019

This study quantified, through a direct harvesting method, storage and annual uptake of carbon from open-grown trees for three landscape tree species frequently planted in the southern region of Korea, and developed quantitative models to easily estimate the carbon reduction by tree growth for each species. The tree species for the study included Camellia japonica, Lagerstroemia indica, and Quercus myrsinaefolia, for which no information on carbon storage and uptake was available. Ten tree individuals for each species (a total of 30 individuals) were sampled considering various stem diameter sizes at given intervals. The study measured biomass for each part of the sample trees to quantify the total carbon storage per tree. Annual carbon uptake per tree was computed by analyzing the radial growth rates of the stem samples at breast height or ground level. Quantitative models were developed using stem diameter as an independent variable to easily calculate storage and annual uptake of carbon per tree for study species. All the quantitative models showed high fitness with $r^2$ values of 0.94-0.98. The storage and annual uptake of carbon from a Q. myrsinaefolia tree with dbh of 10 cm were 24.0 kg and 4.5 kg/yr, respectively. A C. japonica tree and L. indica tree with dg of 10 cm stored 11.2 kg and 8.1 kg of carbon and annually sequestered 2.6 kg and 1.2 kg, respectively. The above-mentioned carbon storage equaled the amount of carbon emitted from the gasoline consumption of about 42 L for Q. myrsinaefolia, 20 L for C. japonica, and 14 L for L. indica. A tree with the diameter size of 10 cm annually offset carbon emissions from gasoline use of approximately 8 L for Q. myrsinaefolia, 5 L for C. japonica, and 2 L for L. indica. The study pioneers in quantifying biomass and carbon reduction for the landscape tree species in the southern region despite difficulties in direct cutting and root digging of the planted trees.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.6
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• pp.1401-1406
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• 2007

The present study investigated neuroprotective effects Rehmanniae Radix on PC12 cells and hippocampal neural cells. PC12 cells were damage by $H_2O_2$ and nitric oxide and organotypic hippocampal slice cultures were damaged by oxygen-glucose deprivation. Then methanol extract of Rehmanniae Radix was treated with 0.5, 5, and $50\;{\mu}g/ml$ in culture media. Effects of Rehmanniae Radix were evaluated with cell viability assay, PI-staining, and TUNEL-labeling. Treatment of Rehmanniae Radix ($with\;5\;and\;50\;{\mu}g/ml$) produced significant increase of cell viability of PC12 cells damaged by $H_2O_2$ and by SNP-induced nitric oxide. Treatment of Rehmanniae Radix produced significant decrease of PI-uptake % in CA1 ($with\;5\;and\;50\;{\mu}g/ml$) and DG ($with\;50\;{\mu}g/ml$) regions of organotypic hippocampal slice cultures damaged by oxygen-glucose deprivation. Moreover, treatment of Rehmanniae Radix produced significant decrease of TUNEL- positive cells in CA1 ($with\;5\;and\;50\;{\mu}g/ml$) and DG ($with\;50\;{\mu}g/ml$) regions of organotypic hippocampal slice cultures damaged by oxygen-glucose deprivation. These results suggest that methanol extract of Rehmanniae Radix has neuroprotective effects on PC12 cells damaged by oxidative stress and on organotypic hippocampal slice cultures damaged by oxygen-glucose deprivation.

• The Journal of Internal Korean Medicine
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• v.25 no.3
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• pp.461-472
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• 2004

Objectives : For the screening of neuroprotective effects of medicinal herbs, the complex system of animal models suffer some disadvantages in controlling critical parameters such as blood pressure and body temperature. Additionally, application of drugs to the appropriate brain area sometimes is difficult, due to poor permeability though the blood brain barrier, and so potential protective effects might be masked. Methods : Organotypic hippocampal slice culture (OHSC) method has the advantages of being relatively easy to prepare and of maintaining the general structure, including tissue integrity and the connections between cells. Drugs can easily be applied and neuronal damage can easily be quantified by using tissues and culture media. This study demonstrates neuroprotective effects of Puerariae radix (葛根, PR), Salviae miltiorrhizae radix (丹蔘, SR), Rhei rhizoma (大黃, RR), and Bupleuri radix (柴胡, BR). These were screenedand compared to MK-801, antagonist of NMDA receptors, by using OHSC of 1 week-old Sprague-Dawley rats. Oxygen/glucose deprivation (OGD) were conducted in an anaerobic chamber $(85%\;N_2,\;10%\;CO_2\;and\;5%\;H_2)$ in a deoxygenated glucose-free medium for 60 minutes. Water extracts of each herbs were treated to culture media with $5\;{\mu}g/ml$ for 48 hours. Results : Neuronal cell death in the cultures was monitored by densitometric measurements of the cellular uptake of propidium iodide (PI). PI fluorescence images were obtained at 48 hours after the OGD and medicinal herb treatment. Also TUNEL-positive cells in the CAI and DG regions and LDH concentrations in culture media were measured at 48 hours after the OGD. According to measured data, MK-801, PR, SR and BR demonstrated significant neuroprotective effect against excessive neuronal cell death and apoptosis induced by the OGD insult. Especially, PR revealed similar neuroprotective effect to MK-801 and RR demonstrated weak neuroprotective effect. Conclusions : These results suggest that OHSC can be a suitable method for screening of neuroprotective effects of medicinal herbs. (This work was supported by the research program of Dongguk University and Grant 01-PJ9-PG1-01CO03-0003 from Ministry of Health & Welfare.)