• Mass Spectrometry Letters
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• v.1 no.1
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• pp.25-28
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• 2010

The high-throughput identification and accurate quantification of proteins are essential strategies for exploring cellular functions and processes in quantitative proteomics. Stable isotope tagging is a key technique in quantitative proteomic research, accompanied by automated tandem mass spectrometry. For the differential proteome analysis of mouse neuronal cell lines, we used a multiplexed isobaric tagging method, in which a four-plex set of amine-reactive isobaric tags are available for peptide derivatization. Using the four-plex set of isobaric tag for relative and absolute quantitation (iTRAQ) reagents, we analyzed the differential proteome in several stroke time pathways (0, 4, and 8 h) after the mouse neuronal cells have been stressed using a glutamate oxidant. In order to obtain a list of the differentially expressed proteins, we selected those proteins which had apparently changed significantly during the stress test. With 95% of the peptides showing only a small variation in quantity before and after the test, we obtained a list of eight up-regulated and four down-regulated proteins for the stroke time pathways. To validate the iTRAQ approach, we studied the use of oxidant stresses for mouse neuronal cell samples that have shown differential proteome in several stroke time pathways (0, 4, and 8 h). Results suggest that histone H1 might be the key protein in the oxidative injury caused by glutamate-induced cytotoxicity in HT22 cells.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.170-175
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• 2012

Clostridium difficile toxin A glucosylates Rho family proteins, resulting in actin filament disaggregation and cell rounding in cultured colonocytes. Given that the cellular toxicity of toxin A is dependent on its receptor binding and subsequent entry into the cell, we herein sought to identify additional colonocyte proteins that might bind to toxin A following its internalization. Our results revealed that toxin A interacted with ERK1 and ERK2 in two human colonocyte cell lines (NCM460 and HT29). A GST-pulldown assay also showed that toxin A can directly bind to ERK1 and ERK2. In NCM460 cells exposed to PMA (an ERK1/2 activator), the phosphorylation of ERK1/2 did not affect the interaction between toxin A and ERK1/2. However, an in vitro kinase assay showed that the direct binding of toxin A to ERK1 or ERK2 inhibited their kinase activities. These results suggest a new molecular mechanism for the cellular toxicity seen in cells exposed to toxin A.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.2
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• pp.121-130
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• 2019

Glutamate toxicity-mediated mitochondrial dysfunction and neuronal cell death are involved in the pathogenesis of several neurodegenerative diseases as well as acute brain ischemia/stroke. In this study, we investigated the neuroprotective mechanism of dieckol (DEK), one of the phlorotannins isolated from the marine brown alga Ecklonia cava, against glutamate toxicity. Primary cortical neurons ($100{\mu}M$, 24 h) and HT22 neurons (5 mM, 12 h) were stimulated with glutamate to induce glutamate toxic condition. The results demonstrated that DEK treatment significantly increased cell viability in a dose-dependent manner ($1-50{\mu}M$) and recovered morphological deterioration in glutamate-stimulated neurons. In addition, DEK strongly attenuated intracellular reactive oxygen species (ROS) levels, mitochondrial overload of $Ca^{2+}$ and ROS, mitochondrial membrane potential (${\Delta}{\Psi}_m$) disruption, adenine triphosphate depletion. DEK showed free radical scavenging activity in the cell-free system. Furthermore, DEK enhanced protein expression of heme oxygenase-1 (HO-1), an important anti-oxidant enzyme, via the nuclear translocation of nuclear factor-like 2 (Nrf2). Taken together, we conclude that DEK exerts neuroprotective activities against glutamate toxicity through its direct free radical scavenging property and the Nrf-2/HO-1 pathway activation.

• Korean Journal of Pharmacognosy
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• v.45 no.3
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• pp.214-219
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• 2014

Artemisia apiacea is a traditional herbal medicine using treatment of eczema and jaundice in Eastern Asia including China, Korea, and Japan. In this study, the three phytosterol constituents were isolated and identified from the hexane fraction of 80% aqueous methanol extract of A. apiacea. Compounds were isolated using open column chromatography (silica gel). Their chemical structures were also established using $^1H$-NMR and $^{13}C$-NMR. Moreover, neuroprotective activity of each compound against glutamate-induced neurotoxicity in hippocampal HT-22 cell line was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Furthermore, Inhibition of reactive oxygen species (ROS) and calcium ion ($Ca^{2+}$) accumulation were measured for elucidation of neuroprotective mechanism of isolated compounds. They showed that stigmasterol had neuroprotective activity against the glutamate-induced toxicity by inhibition of ROS and $Ca^{2+}$ production. In conclusion, isolated compound of A. apiacea might be useful for therapeutic agent against neurodegenerative diseases.

• Fisheries and Aquatic Sciences
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• v.22 no.5
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• pp.9.1-9.7
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• 2019

The study investigated the changes in growth performance, hematological parameters, plasma components, and stress indicators of juvenile starry flounder, Platichthys stellatus, depending on varying exposure to hexavalent chromium. P. stellatus was exposed to waterborne chromium at 0, 50, 100, 200, and 400 ppb for 4 weeks. The result showed that Cr exposure resulted in decreased daily length gain (DLG), daily weight gain (DWG), condition factor (CF), and hepatosomatic index (HIS) in P. stellatus. In terms of hematological parameters, red blood cell (RBC) count, hematocrit (Ht), and hemoglobin (Hb) significantly decreased at 400 ppb after 2 weeks. In terms of plasma components, inorganic analysis was unchanged and cholesterol, an organic component, considerably increased at 400 ppb after 4 weeks. Plasma enzyme components including glutamic oxalate transaminase (GOT) and glutamic pyruvate transaminase (GPT) were significantly increased. Stress indicators such as cortisol and glucose were notably increased over 100 ppb after 4 weeks with increasing chromium concentration. The results indicate that exposure to waterborne Cr induced toxic effects on growth, hematological parameters, plasma components, and stress indicators.

• Journal of Life Science
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• v.25 no.6
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• pp.656-662
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• 2015

Natural products and natural product structures play a general and highly significant role in drug discovery and development process because it has various merits and potentials for new drug source that have extensive clinical experience, development time contraction, excellent stability and safety. In several neurological disorders, neuronal death and excessive activation of microglia (neuro-inflammation) are observed. A number of drug discovery-related neuronal cell death and neuro-inflammation was studied from natural products, respectively. However, until now, it has not been possible to study dual-protection molecules recorded in the Natural Product library. In the present study, using the natural product-derived library of the Institute for Korea Traditional Medical Industry, we investigated dual-protective molecules against glutamate (a classical excitatory neurotransmitter)-induced oxidative stress mediated neuronal cell death and LPS-induced excessive activated microglial cells (immune cells of the brain). Chrysophanol, extracted from Rheum palmatum, had dual-protective effects against both glutamate-induced neuronal cell death and LPS-induced NO production, triggering proinflammatory cytokines and microglia activation and resulting in neuroinflammation. Flow-cytometry analysis revealed that chrysophanol had a scavenger effect, scavenging glutamate- and LPS-induced reactive oxygen species (ROS) produced by neuronal and microglial cells, respectively. Based on the present study, chrysophanol may have an important protective role against neuronal cell death and neuroinflammation in the brain. The results may be helpful for studying drug development candidates for treating central nervous system disorders.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.465-472
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• 2001

The effects of water level reduction in rearing tank and fish transference during fish selection process on the stress response (hematological factors, cortisol, glucose, lactic acid and osmolality) of tank-reared olive flounder Paralichthys olivaceus of large (FL), small (FS) and Japanese croaker, Nibea japonica (JC) were examined in running seawater culture system. The water level of rearing unit was lowered from 33 cm to 8 cm in the course of 2 minutes in the water level reduction experiment. The fish were removed from rearing tank (12 ton) to 450 L tank in 30 seconds after capture in the fish transference experiment, In water level reduction, the hematocrit of FL was significantly increased from $14.6\%$ at beginning to $23.5\%$ after 10 hours, However, it decreased to the value of beginning after 46 hours. Plasma cortisol concentration of FL was the highest concentration (13.7 ng/mL) after 22 hours, but it decreased to 4.0 ng/mL at the end of experiment. Cortisol concentration of FS did not show any significant difference during the experiment. The cortisol concentration of JC were significantly higher at 4 hours (282.3 ng/mL) and 22 hours (350.5 ng/mL), Glucose concentration of JC was the highest (138.0 mg/dL) at 22 hours. Lactic acid concentration was not different between experimental groups. In the fish transference experiment, red blood cell of FL was increased from $1.9\times10^6\;cell/{\mu}\;L\;to\;4.2\times10^6\;cell/{\mu}L$ in 24 hours. Blood hemoglobin of JC were significantly elevated in 24 hours. At 1 hour after transference, plasma cortisol concentrations in both fish species were increased to 95.3 ng/mL in FL and 175.5 ng/mL in JC. Glucose concentration of JC was increased to 132.5 mg/dL at 1 hour, 129.5 mg/dL at 3 hours after transference.

• Korean Journal of Pharmacognosy
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• v.45 no.1
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• pp.48-54
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• 2014

Alzheimer's disease is progressive neurodegenerative disorder by the loss of memory and learning abilities. Codonopsis lanceolata (C. lanceolata) is traditional medicinal plant used for the treatment of inflammatory diseases. The aim of study was to evaluate the effect of steamed C. lanceolata on scopolamine-induced memory impairment in the Morris water maze test and passive avoidance test. In addition, this study investigated the neuroprotective effects of steamed C. lanceolata on glutamate-induced cell death in HT22 cells using MTT assay. The results showed that steamed C. lanceolata (500 mg/kg body weight, p.o.) reversed spatial memory impairment by scopolamine in Morris water maze test and passive avoidance test. Steamed C. lanceolata attenuated memory impairment by scopolamine compared with common C. lanceolata. In addition, administration of steamed C. lanceolata significantly also reduced cell death. We suggest that steaming process more improve cognitive enhancing and neuroprotective effect of C. lanceolata than common C. lanceolata.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.13
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• pp.5371-5376
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• 2015

Berberine (B1), isolated from stems of Coscinium fenestratum (Goetgh.) Colebr, was used as a principle structure to synthesize three phenolic derivatives: berberrubine (B2) with a single phenolic group, berberrubine chloride (B3) as a chloride counter ion derivative, and 2,3,9,10-tetra-hydroxyberberine chloride (B4) with four phenolic groups, to investigate their direct and indirect antioxidant activities. For DPPH assay, compounds B4, B3, and B2 showed good direct antioxidant activity ($IC_{50}$ values=$10.7{\pm}1.76$, $55.2{\pm}2.24$, and $87.4{\pm}6.65{\mu}M$, respectively) whereas the $IC_{50}$ value of berberine was higher than $500{\mu}M$. Moreover, compound B4 exhibited a better DPPH scavenging activity than BHT as a standard antioxidant ($IC_{50}=72.7{\pm}7.22{\mu}M$) due to the ortho position of hydroxyl groups and its capacity to undergo intramolecular hydrogen bonding. For cytotoxicity assay against human fibrosarcoma cells (HT1080) using MTT reagent, the sequence of $IC_{50}$ value at 7-day treatment stated that B1 < B4 < B2 ($0.44{\pm}0.03$, $2.88{\pm}0.23$, and $6.05{\pm}0.64{\mu}M$, respectively). Berberine derivatives, B2 and B4, showed approximately the same level of CAT expression and significant up-regulation of SOD expression in a dose-dependent manner compared to berberine treatment for 7-day exposure using reverse transcription-polymerase chain reaction (RT-PCR) assays. Our findings show a better direct-antioxidant activity of the derivatives containing phenolic groups than berberine in a cell-free system. For cell-based system, berberine was able to exert better cytotoxic activity than its derivatives. Berberine derivatives containing a single and four phenolic groups showed improved up-regulation of SOD gene expression. Cytotoxic action might not be the main effect of berberine derivatives. Other pharmacological targets of these derivatives should be further investigated to confirm the medical benefit of phenolic groups introduced into the berberine molecule.

• YAKHAK HOEJI
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• v.58 no.5
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• pp.287-293
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• 2014

Alzheimer's disease (AD), most common form of dementia is characterized that memory deficit and loss of cognitive function. This study was evaluated cognitive enhancing effect of water and ethanol extracts of Codonopsis lanceolata and compared using Morris water maze and passive avoidance test. The water and 70% ethanol extracts (100, 300 and 500 mg/kg) were administered to mice. The neuroprotective effect on glutamate-induced cell death in HT22 cells was additionally investigated using MTT assay. Results showed 70% ethanol extract of Codonopsis lanceolata enhanced cognitive function than water extract, as shown by decrease in escape latency time in Morris water maze test. In passive avoidance test, 70% ethanol extract also increased the latency time compared to the water extract. Furthermore, 70% ethanol extract significantly protected neuronal cell against glutamate cytotoxicity and showed higher than neuroprotective effect of water extract. These results indicate that 70% ethanol extract more improve spatial cognitive ability and protected neuronal cells than water extract.