• The Journal of Internal Korean Medicine
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• v.40 no.3
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• pp.425-442
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• 2019

Objectives: This study aimed to reveal the pharmacological properties of the newly prescribed herbal mixture, Chenmadansamgamibokhap-tang(CDBT), against hypoxia-induced neuronal cell injury (especially mouse hippocampal neuronal cell line, HT-22 cells) and their corresponding mechanisms. Methods: A cell-based in vitro experiment, in which a hypoxia condition induced neuronal cell death, was performed. Various concentrations of the CDBT were pre-treated to the HT-22 cells for 4 h before 18 h in the hypoxia chamber. The glial cell BV-2 cells were stimulated with $IFN{\gamma}$ and LSP to produce inflammatory cytokines and reactive oxygen species. When the neuronal HT-22 cells were treated with this culture solution, the drug efficacy against neuronal cell death was examined. Results: CDBT showed cytotoxicity in the normal condition of HT-22 cells at a dose of $125{\mu}g/mL$ and showed a protective effect against hypoxia-induced neuronal cell death at a dose of $31.3{\mu}g/mL$. CDBT prevented hypoxia-induced neuronal cell death in a dose-dependent manner in the HT-22 cells by regulating $HIF1{\alpha}$ and cell death signaling. CDBT prevented neuronal cell death signals and DNA fragmentation due to the hypoxia condition. CDBT significantly reduced cellular oxidation, cell death signals, and caspase-3 activities due to microglial cell activations. Moreover, CDBT significantly ameliorated LPS-induced BV-2 cell activation and evoked cellular oxidation through the recovery of redox homeostasis. Conclusions: CDBT cam be considered as a vital therapeutic agent against neuronal cell deaths. Further studies are required to reveal the other functions of CDBT in vivo or in the clinical field.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.5
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• pp.1019-1024
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• 2009

Diabetic neuropathy is characterized by the decrease of cell viability in neuron, which is induced by the hyperglycemia. HT22 cell is the neuron cell line originated from hippocampus. Ginsenosides have been reported to retain anti-diabetic effect. However, the preventive effect of ginsenosides in the condition of diabetic neuropathy was not elucidated. Thus, this study was conducted to examine the protective effect of ginsenoside total saponin (GTS), panoxadiol (PD), and panoxatriol (PT) in the high glucose-induced cell death of HT22 cells, an in vitro cellular model for diabetic neuropathy. In present study, high glucose increased lactate dehydrogenase(LDH) activity, the lipid peroxide(LPO) formation and induced the decrease of cell viability. These effects were completely prevented by the treatment of GTS, but partially prevented by the treatment of PD and PT. High glucose also increased the expression of Bax and cleaved form of caspase-3 but decreased that of Bcl-2. These effects of high glucose on Bax, Bcl-2 and cleaved form of caspase-3 were completely prevented by the treatment of GTS, but partially prevented by the treatment of PD and PT in HT22 cells. In conclusion, ginsenosides prevented high glucose-induced cell death of hippocampal neuron through the inhibition of oxidative stress and apoptosis in HT 22 cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.886-890
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• 2017

In this study, the protective effect of rice with Phellinus linteus mycelium (PLMR) against hydrogen peroxide-induced oxidative stress was assessed in a mouse hippocampal neuronal HT22 cell line through (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) salt (MTS) assay and western blot. MTS assay using HT22 cells showed that PLMR extract did not affect viability at a concentration range from 1 mg/mL to 5 mg/mL. However, at concentrations over 10 mg/mL, PLMR extract resulted in increased cell death. Cell viability of HT22 was significantly reduced by $H_2O_2$ treatment, and reduction of cell viability was efficiently restored by treatment with PLMR extract in a dose-dependent manner from 0.1 to 1 mg/mL. Cells treated with $H_2O_2$ showed increased expression of Bax, a pro-apoptotic protein, which was down-regulated by treatment with PLMR extract. On the other hand, cells treated with $H_2O_2$ resulted in reduced expression of Bcl-2, an anti-apoptotic protein, which was restored by treatment with PLMR extract. In addition, treatment with PLMR extract reduced expression of cleaved caspase 3 and PARP, which were up-regulated by $H_2O_2$ treatment. The results may suggest that treatment with PLMR extract would suppress $H_2O_2$-induced apoptosis of HT22 cells.

• Natural Product Sciences
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• v.15 no.1
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• pp.32-36
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• 2009

Glutamate causes neurotoxicity through formation of reactive oxygen species and activation of mitogen-activated protein kinase (MAPK) pathways. MAPK phosphatase-1 (MKP-1) is one of the phosphatases responsible for dephosphorylation/deactivation of three MAPK families: the extracellular signal-regulated kinase-1/2 (ERK-1/2), the c-Jun N-terminal kinase-1/2 (JNK-1/2), and the p38 MAPK. In this report, the potential involvement of MKP-1 in neuroprotective effects of curcumin, the active ingredient of turmeric (Curcuma longa), was examined using HT22 cells. Glutamate caused cell death and activation of ERK-1/2 but not p38 MAPK or JNK-1/2. Blockage of ERK-1/2 by its inhibitor protected HT22 cells against glutamate-induced toxicity. Curcumin attenuated glutamate-induced cell death and ERK-1/2 activation. Interestingly, curcumin induced MKP-1 activation. In HT22 cells transiently transfected with small interfering RNA against MKP-1, curcumin failed to inhibit glutamate-induced ERK-1/2 activation and to protect HT22 cells from glutamate-induced toxicity. These results suggest that curcumin can attenuate glutamate-induced neurotoxicity by activating MKP-1 which acts as the negative regulator of ERK-1/2. This novel pathway may contribute to and explain at least one of the neuroprotective actions of curcumin.

• Korean Journal of Pharmacognosy
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• v.39 no.3
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• pp.213-217
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• 2008

Oxidative stress is considered to play an important role in a variety of neurodegenerative disorders of central nervous system. The immortalized mouse hippocampal cell line, HT22, phenotypically resembles neuronal precursor cells but lacks functional ionotropic glutamate receptors, thus excluding excitotoxicity as a cause for glutamate triggered cell death. Therefore, HT22 cells are a useful model for studying oxidative glutamate toxicity. In this study, we examined whether the methanol extracts of some native plants at Mt. Baekdu could protect HT22-immortalized hippocampal cells against glutamate-induced oxidative stress. Seventy-eight plants sources were collected at Mt. Baekdu, and extracted with methanol. These extracts had been screened the protective effects against glutamate-induced oxidative damage in HT22 cells at the 100 and 300 ${\mu}g/ml$. Of these, thirteen methanolic extracts, Acer mono (leaf), Artemisia stolonifera (aerial part), Carduus crispus (aerial part), Carex mongolica (whole plant), Clematis hexapetala (whole plant), Galeopsis bifida (aerial part), Galium verum (whole plant), Ganoderma lucidum (whole plant), Ixeris chinensis (whole plant), Malva verticillata (aerial part), Polygonum senticosum (whole plant), Rebes mandshricum (branch), and Taraxacum mongolicum (aerial part), showed significant protective effects against glutamate-induced oxidative damage in HT22 cells.

• Journal of Life Science
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• v.26 no.12
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• pp.1458-1465
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• 2016

We investigated the neuroprotective effect of an ethanol extract from Asparagus cochinchinensis (AC) against glutamate-induced toxicity in the HT22 hippocampal cell, which is an ideal in vitro model for oxidative stress. The neuroprotective effects of AC in HT22 cells were evaluated by analyzing cell viability, lactate dehydrogenase (LDH), flow cytometry for cell death types, reactive oxygen species (ROS), mitochondria membrane potential (MMP), and Western blot assays. In the cell death analysis, AC treatment resulted in significantly attenuated glutamate-induced loss of cell viability with a decrease in LDH release. AC treatment also reduced glutamate-induced apoptotic cell death. In the ROS and MMP analysis, AC treatment inhibited the elevation of intracellular ROS induced by glutamate exposure and the disruption of MMP. In oxidative stress-related proteins analysis, AC treatment inhibited the expression of poly ADP ribose polymerase and heme oxygenase-1 by glutamate. These results indicate that AC exerts a significant neuroprotective effect against glutamate-induced hippocampal damage by decreasing ROS production and stabilizing MMP. Thus, AC potentially provides a new strategy for the treatment of oxidative stress-related diseases.

• Korean Journal of Pharmacognosy
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• v.53 no.2
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• pp.79-86
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• 2022

Excessive glutamate causes oxidative stress in neuronal cells, which can cause degenerative neurological disorders. We tried to find medicinal plant showed neuroprotective activity by using glutamate-injured HT22 cell as a model system. In this study, we found that Boesenbergia rotunda methanol extract showed neuroprotective activity against glutamate induced neurotoxicity in mouse hippocampal HT22 cells. B. rotunda methanol extract suppressed the formation of reactive oxygen species and decreased intracellular Ca²⁺concentration. Also, B. rotunda made mitochondrial membrane potential maintain to normal levels. In addition, B. rotunda increased total glutathione amount and activated antioxidative enzyme such as glutathione reductase and glutathione peroxidase compared to glutamate-treated groups. These results suggested that B. rotunda decreased neuronal cell death damaged by high concentrations of glutamate treatment, via antioxidative mechanism and might be one of candidate of development of new drug to treat neurodegenerative disease such as Alzheimer's disease.

• Korean Journal of Pharmacognosy
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• v.43 no.4
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• pp.316-322
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• 2012

Siegesbeckia Herba is known to have anti-oxidant, anti-inflammatory, anti-allergic and anti-tumor. The objective of this study is to explore the neuroprotective effect of Siegesbeckia Herba against glutamate-induced oxidative stress in mouse hippocampal HT22 cells. Siegesbeckia Herba 70% ethanol extract and solvent fractions have the potent neroprotective effects on glutamate-induced nerotoxicity by induced the expression of heme oxygenase (HO)-1 in the mouse hippocampal HT22 cells. Especially, ethyl acetate fraction showed higher protective effect. In HT22 cell, Siegesbeckia Herba ethyl acetate fraction makes the nuclear accumulation of Nrf2. Further, we found that treatment with c-JUN N-terminal kinase (JNK) inhibitor (SP600125) reduced Siegesbeckia Herba ethyl acetate fraction induced HO-1 expression and Siegesbeckia Herba ethyl acetate fraction also increased JNK phosphorylation. In conclusion, the ethyl acetate fraction of 70% ethanol extract of Siegesbeckia Herba significantly protect glutamate-induced oxidative damage by induction of HO-1 via Nrf2 and JNK pathway in mouse hippocampal HT22. Taken together these finding suggest that Siegesbeckia Herba ethyl acetate fraction good source for taking active compounds and may be a potential therapeutic for brain disorder by targeting the oxidative stress of neuronal cell.

• The Journal of Korean Medicine
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• v.39 no.1
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• pp.35-43
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• 2018

Objectives: In the brain, glutamate is the most important excitable neurotransmitter in physiological and pathological conditions. However, the high level of glutamate induces neuronal cell death due to exitotoxicity and oxidative stress. The present study investigated to evaluate a possible neuroprotective effect of furosin isolated from Euphorbia helioscopia against glutamate-induced HT22 cell death. Methods: Furosin was isolated from methanol extract of Euphorbia helioscopia and examined whether it protects glutamate-induced neuronal cell death. The cell viability was determined using Ez-Cytox assay. Anti-oxidative effect of furosin was determined by DPPH scavenging activities, and the levels of intracellular reactive oxygen species (ROS) were determined by the fluorescent intensity after staining the cells with $H_2DCFDA$. To evaluate apoptotic cell death, we performed nuclear staining and image-based cytometeric analysis. Results: The cell viability was significantly increased by treatement with furosin compared with the treatment with glutamate. Furosin showed a strong DPPH radical scavenging activity ($EC50=1.83{\mu}M$) and prevented the accumulation of intra cellular ROS. Finally, the presence of 50 and $100{\mu}M$ furosin significantly the percentage of apoptotic cells compared with glutamate treatment. Conclusion: The present study found that furosin is a potent neuroprotectant against glutamate-induced oxidative stress through inhibition of apoptotic cell death induced by glutamate. Therefore, the present study suggests that furosin as a bioactive compound of E. helioscopia can be a useful source to develop a drug for the treatment of neurodegenerative diseases and acute brain injuries.

• YAKHAK HOEJI
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• v.47 no.2
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• pp.85-92
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• 2003

Seok-jeong (SJ) is a solution of various metal ions and numerous other organic substances produced through extraction and fermentation of herbs and soil using geo-microbes, and it has been shown to improve symptoms of senile dementia. In this study, we investigated the protective effects of SJ against neurotoxicity of cadmium in HT22 hippocampal neuron cell line. SJ significantly protected from the cadmium-induced decreased cell viability measured by MTT assay (p＜0.01). The protective effects of SJ against cadmium toxicity were confirmed through observing morphological changes using inverted microscope. Additionally, SJ significantly repressed the formation of lipid peroxidation induced by high concentration of cadmium, and likewise, significantly repressed the reduction of glutathione by cadmium in HT22 cells. Vitamin C at the concentration found in SJ did not show any protective effect against cadmium toxicity in HT22 cells, indicating that vitamin C may not have a major role in the protective mechanism of SJ. Taken together, these results suggest that SJ may be a valuable agent for the protection of cadmium toxicity on the neuronal cells, and that the mechanism of the action of SJ may be due to reduced lipid peroxidation and increased glutathione level.