• Herbal Formula Science
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• v.22 no.2
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• pp.35-43
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• 2014

Objectives : The purpose of this study was to identify antiproliferative effects of Salviae miltiorrhizae Radix(SM) extracts against cancer cell lines. Methods : We used 2 kinds of cancer cell lines such as colon cancer cells(HT-29), human oral epitheloid carcinoma cells(KB). MTT assay was performed to examine the efficacy of SM extracts on the cytostaticity of cancer cells in proportion to time and doses. Apoptosis was evaluated by DNA laddering and DAPI nuclei staining. Results : The MTT absorbances against HT-29 and KB of SM extracts were significantly decresed. DNA ladders could be identified in KB of SM extracts. The morphological change were observed and number of cells were decreased by SM extracts. Conclusions : SM extracts is considered to be effective to induce apoptosis and inhibit cancer cell proliferation.

• Natural Product Sciences
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• v.29 no.2
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• pp.67-73
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• 2023

Oxidative stress brings about apoptosis through various mechanisms. In particular, oxidative stress in neuronal cells can causes a variety of brain diseases. This study was conducted to investigate the effect of Bidens tripartita on oxidative stress in neuronal cells. B. tripartita has traditionally been used in Russia as a medicine for diseases such as rhinitis, angina and colitis. Over-production of glutamate induces oxidative stress. When the oxidative stress occurs in the cells, reactive oxygen species (ROS) and Ca²⁺ increase. In addition, the abrupt decline of mitochondrial membrane potential and the decrease of glutathione related enzymes such as glutathione reductase (GR) and glutathione peroxidase (GPx) are also observed. The samples used in the experiment showed cytoprotective effect in the MTT assay. It also lowered the ROS and Ca²⁺ level, and increased degree of mitochondrial membrane potential, GR and GPx. As a result, B. tripartita had a positive effect against oxidative stress. Thus, it is expected to have potential for treatment and prevention of degenerative brain diseases such as Alzheimer's disease.

• Journal of Life Science
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• v.32 no.1
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• pp.11-22
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• 2022

N-methyl-D-aspartate (NMDA) receptors have received considerable attention regarding their involvement in glutamate-induced neuronal excitotoxicity. Resveratrol has been shown to exhibit neuroprotective effects against this kind of overactivation, but the underlying cellular mechanisms are not yet clearly understood. In this study, HT-22 neuronal cells were treated with NMDA in Mg²⁺-free buffer and subsequently used as an experimental model of glutamate excitotoxicity to elucidate the mechanisms of resveratrol-induced neuroprotection. We found that NMDA treatment causes a drop in MTT reduction ability, disrupts inside-negative transmembrane potential of mitochondria, depletes cellular ATP levels, and stimulates intracellular ROS production. Double fluorescence imaging studies demonstrated an increased formation of mitochondrial permeability transition (MPT) pores accompanied by apoptotic cell death, while cobalt protoporphyrin and bilirubin showed protective effects against NMDA-induced mitochondrial injury. On the other hand, zinc protoporphyrin IX significantly attenuated the protective effects of resveratrol which was itself shown to enhance heme oxygenase-1 (HO-1) mRNA and protein expression levels. In cells transfected with HO-1 small interfering RNA, resveratrol failed to suppress the NMDA-induced effects on MTT reduction ability and MPT pore formation. The present study suggests that resveratrol may prevent mitochondrial injury in NMDA- treated HT-22 cells and that enhanced expression of HO-1 is involved in the underlying cellular mechanism.

• Archives of Pharmacal Research
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• v.28 no.7
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• pp.804-809
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• 2005

The EtOAc-soluble fraction from a methanolic extract of Hovenia dulcis Thunb. exhibited neuroprotective activity against glutamate-induced neurotoxicity in mouse hippocampal HT22 cells. The neuroprotective activity-guided isolation resulted in 8 phenolic compounds (1-8), such as vanillic acid (1), ferulic acid (2), 3,5-dihydroxystilbene (3), (+)-aromadendrin (4), methyl vanillate (5), (-)-catechin (6), 2,3,4-trihydrobenzoic acid (7), and (+)-afzelechin (8). Among these, compounds 6 and 8 had a neuroprotective effect on the glutamate-induced neurotoxicity in HT22 cells. Furthermore, compound 6 had a DPPH free radical scavenging effect with an $IC_{50}$ value of $57.7{\mu}M$, and a superoxide anion radical scavenging effect with an $IC_{50}$ value of $8.0{\mu}M$. Both compounds 6 and 8 had ABTS cation radical scavenging effects with $IC_{50}$ values of $7.8{\mu}M\;and\;23.7${\mu}M$, respectively. These results suggest that compounds 6 and 8 could be neuroprotectants owing to their free radical scavenging activities.

• The Journal of Internal Korean Medicine
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• v.22 no.1
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• pp.79-85
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• 2001

Object : Hepatitis Treatment-tang No.1 has been used for the treatment of Liver disease and Jaundice. Long-term EtOH exposure leads to immunoregulatory and detoxification impairment. This study aimed to determine the relationship between TNF-${\alpha}$ production and expression, and EtOH-induced cytotoxicity on Hep G2 cells. Method : Cells were incubated with EtOH in the presence or absence of HT. The cells were tested after 24 hours and, again, after 48 hours. Cytoviability and TNF-${\alpha}$ release were analyzed by MTT assay and enzyme linked immunosorbent assay (ELISA), respectively. After 24 hours of EtOH exposure, the cytoviability decreased, and the release of TNF-${\alpha}$ was increased. Increased amounts of TNF-${\alpha}$ contribute to EtOH-induced cytotoxicity. The Anti-TNF-${\alpha}$ antibody almost abolished it. Interestingly, EtOH-induced cytotoxicity and TNF-${\alpha}$ production were inhibited by HT. Moreover, when HT was used in combination with the anti-TNF-${\alpha}$ antibody, there was a marked inhibition of EtOH-induced cytotoxicity. Results : These results suggest that HT may prevent the cytotoxicity through partial inhibition of the TNF-${\alpha}$ secretion.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.2
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• pp.224-230
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• 2014

Marine algae have long been recognized as a health and beauty food, based on its anti-tumor, anti-inflammatory and anti-obesity activities. In this study, methanol extracts were prepared from 10 different phaeophyta, after which DPPH radical scavenging and cytoprotective activities of HT-22 cells against ${\beta}$-amyloid protein ($A{\beta}$), which has neurotoxic effects, were investigated. In DPPH experiments, Ecklonia cava and Ishige okamurai showed strong ROS scavenging activities, whereas eight other phaeophyta including Petalonia binghamiae (P. bin) showed weak ROS scavenging activities. To validate the cytoprotective effects of 10 different phaeophyta in $A{\beta}$-induced HT-22 cells, protein expression levels of APP, BACE1, iNOS, phosphorylated ERK1/2, phosphorylated p38 and phosphorylated JNK1/2 were determined along with MTT assay. In the MTT assay, P. bin showed the best effective cytoprotective activity at a concentrations of $25{\mu}g/mL$, whereas Sargassum confusum, Colpomenia sinuosa, Myelophycus simplex, and Sargassum hemiphyllum showed potential. Determination of protein expression levels related to $A{\beta}$-induced neurotoxicity in the five selected phaeophyta showed that P. bin inhibited BACE1 and iNOS expression in $A{\beta}$-induced HT-22 cells. These results indicate that the cytoprotective effects of P. bin are mediated by suppressing the pathways involving $A{\beta}$-induced ERK and p38 activation.

• BMB Reports
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• v.49 no.11
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• pp.617-622
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• 2016

Oxidative stress is closely associated with various diseases and is considered to be a major factor in ischemia. NAD(P)H: quinone oxidoreductase 1 (NQO1) protein is a known antioxidant protein that plays a protective role in various cells against oxidative stress. We therefore investigated the effects of cell permeable Tat-NQO1 protein on hippocampal HT-22 cells, and in an animal ischemia model. The Tat-NQO1 protein transduced into HT-22 cells, and significantly inhibited against hydrogen peroxide ($H_2O_2$)-induced cell death and cellular toxicities. Tat-NQO1 protein inhibited the Akt and mitogen activated protein kinases (MAPK) activation as well as caspase-3 expression levels, in $H_2O_2$ exposed HT-22 cells. Moreover, Tat-NQO1 protein transduced into the CA1 region of the hippocampus of the animal brain and drastically protected against ischemic injury. Our results indicate that Tat-NQO1 protein exerts protection against neuronal cell death induced by oxidative stress, suggesting that Tat-NQO1 protein may potentially provide a therapeutic agent for neuronal diseases.

• Journal of Microbiology and Biotechnology
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• v.28 no.2
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• pp.246-254
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• 2018

Enzyme fermentation is a type of food processing technique generally used to improve the biological activities of food and herbal medicines. In this study, a Syzygii Flos (clove) extract was fermented using laccase derived from Trametes versicolor (LTV). The fermented clove extract showed greater neuroprotective effects against glutamate toxicity on HT22 than the non-fermented extract did. HPLC analysis revealed that the eugenol (1) and dehydrodieugenol (2) contents had decreased and increased, respectively, after fermentation. The content of 2 peaked at 1 h after fermentation to $103.50{\pm}8.20mg/g_{ex}$ (not detected at zero time), while that of 1 decreased to $79.54{\pm}4.77mg/g_{ex}$ ($185.41{\pm}10.16mg/g_{ex}$ at zero time). Compound 2 demonstrated promising HT22 neuroprotective properties with inhibition of $Ca^{2+}$ influx, the overproduction of intracellular reactive oxygen species, and lipid peroxidation. In addition, LTV showed the best fermentation efficacy compared with laccases derived from Pleurotus ostreatus and Rhus vernicifera.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.5
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• pp.1334-1336
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• 2006

Effects of twenty-three aqueous herbal extracts used in oriental medicines on heme oxygenase (HO)-1 expression were estimated in a mouse hippocampal cell line, HT22. HO-1 is one of the cytoprotective enzymes activated various stimuli, and Western blot analysis was used for evaluated HO-1 expression. Six aqueous extracts such as Rhei Rhizoma, Paeoniae Radix, Uncariae Ramulus et Uncus, Theae Folium, Prunellae Spica, and Coptidis Rhizoma significantly increased HO-1 expression in HT22 cells at the concentration of 300 ${\mu}$g/ml. In Addition, four aqueous extracts including Eucommiae Cortex, Moutan Cortex Radicis, Ginseng Radix Rubra, and Scutellariae Radix moderately increased HO-1 expression. These results would be usefulfor the isolation and identification of their neuroprotective principles.

• The Korean Journal of Physiology
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• v.22 no.2
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• pp.245-257
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• 1988

The present study was undertaken to investigate the effect of clonidine on the response of the dorsal horn cells to intra-arterially administered bradykinin $(BK:40{\mu}g)$ and $K^+(4mg)$ in spinal cats and cats with intact spinal cord. The change in the activities of low threshold (LT), high threshold (HT) and wide dynamic range (WDR) cells induced by BK and $K^+$ were determined before and after treatment of animals with clonidine. Also studied was mechanism of inhibitory action of clonidine on the responses of dorsal horn cells to the chemical algogenics. Number of WDR cell responded to intra-arterially administered BK and $K^+$ was greater in spinal animals than in cats with intact spinal cord. Following administration of BK or $K^+$ no change was observed in the activity of LT cell whereas activity of HT cell increased invariably. The increased response of HT cell to BK and $K^+$ was markedly suppressed by clonidine. On the other hand, such inhibitory actions of clonidine were almost completely blocked by yohimbine. The majority of WDR cells were activated by $K^+$ while response of WDR cells to BK was diverse (excitatory, inhibitory or mixed). These results indicate that clonidine inhibits responses of the dorsal horn cells not only to thermal or mechanical stimulations but also to chemical algogenics, and that the inhibitory action of clonidine is generally mediated through excitation of ${\alpha}_2-adrenoreceptors$.