• Journal of Oriental Neuropsychiatry
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• v.20 no.1
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• pp.59-73
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• 2009

Objectives : This experiment was designed to investigate the effect of the ODTCMT hot water extract & ultra-fine Powder on Alzheimer's Disease Model Induced by 13A. Methods : The effects of the ODTCMT hot water extract on expression of IL-l${\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. The effects of the ODTCMT hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine were investigated. Results : 1. The ODTCMT hot water extract significantly suppressed the production of TNF-${\alpha}$, IL-6 and IL-l${\beta}$ in BV2 microglial cell line treated with LPS. 2. The ODTCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine-induced impairment of memory in the experiment of Morris water maze. Conclusions : These results suggest that the ODTCMT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the ODTCMT hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.

• Natural Product Sciences
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• v.18 no.3
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• pp.183-189
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• 2012

It is well known that inflammation is associated with neurodegenerative disorders, including Alzheimer' disease, Parkinson's disease and ischemia. Nitric oxide (NO), a pro-inflammatory mediator, is produced by inducible NO synthase (iNOS) in microglia as well as macrophages and appears to account for neurodegeneration. In this study, we aimed to isolate NO inhibitors from Pyrus pyrifolia by activity guided purification. As a result, we identified daucosterol and ${\beta}$-sitosterol, which have not been isolated from this plant before. This article also describes NO inhibitory activities of the methanol extract of Pyrus pyrifolia fruit and the isolated compounds from this, which are lupeol, betulin, betulinic acid, ${\beta}$-sitosterol and daucosterol, in LPS-activated RAW 264.7 and BV2 cell lines. Western blot analysis was performed to clarify the underlying mechanism of NO inhibition in the two cell lines.

• International Journal of Industrial Entomology and Biomaterials
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• v.29 no.2
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• pp.207-213
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• 2014

The structural proteins of classical swine fever virus (CSFV) consist of nucleocapsid protein C and envelope glycoprotein $E^{rns}$ (E0), E1 and E2. Among them, E2, the most immunogenic of the CSFV glycoproteins, induces a protective immune response in swine. In this study, to determine the optimal expression conditions of glycoprotein E2 using baculovirus system, we investigated the influence of insect cells and media to the expression of recombinant E2. Recombinant virus containing glycoprotein E2 coding gene was constructed with bApGOZA DNA. Expression of the glycoprotein E2 was analyzed by SDS-PAGE and Western blot analysis using anti-CSFV E2 monoclonal antibodies. Expression of glycoprotein E2 in Sf21 cells was first observed after 3 days and reached a maximum on the 5th day after infection. Furthermore, the highest levels of glycoprotein E2 expression were observed at multiplicity of infection (MOI) of 5. When three different insect cell lines (Sf21, High-Five and Se301) were tested, High-Five cells showed the highest production. In addition, four different serum-free and serum-supplemented media, respectively, were tested for the expression of glycoprotein E2 and the budded virus (BV) titers. As a result, serum-supplemented medium provided the best conditions for protein production and the BV yield.

• Journal of Acupuncture Research
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• v.22 no.2
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• pp.147-154
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• 2005

Objective: In this study, the effects of Liriope Platyphylla against LPS-induced inflammation was investigated. Methods: Cell viability was determined using the MTT assay. To identify expressions of COX-2 and iNOS mRNA, RT-PCR was performed. Assessment of PGE2 synthesis was performed using the PGE2 immunoassay. Measurement of NO synthesis was performed using the NO detection. Result : The MTT assay revealed that Liriope Platyphylla exerted no significant cytotoxicity in the microglial BV2 cells. RT-PCR analysis revealed that the mRNA levels of COX-2 and iNOS were significantly decreased in the LPS- and 5 mg/ml Liriope Platyphylla treated group. From the PGE2 immunoassay and NO detection, PGE2 and NO synthesis was significantly suppressed in the LPS- and 5 mg/ml Liriope Platyphylla treated group. Conclusion : In these study, Liriope Platyphylla was shown to suppress PGE2 and NO production by inhibiting LPS-stimulated enhancement of COX-2 enzyme activity and iNOS expression. It is very possible that Liriope Platyphylla can offer a valuable mode of therapy for the treatment of brain inflammatory diseases.

• Journal of Oriental Neuropsychiatry
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• v.18 no.2
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• pp.101-132
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• 2007

Objective : This research investigates the effect of the NogYongDaeBoTang,(NYDBT) on Alzheimer's disease. Method : The effects of the NYDBT extract on (1) $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) acetylcholinesterase(AChE), amyloid precursor proteins(APP), and glial fibrillary acidic protein(GFAP) mRNA the AChE activity and the APP production of PC-12 cell treated with CT-105; (3) the behavior; (4) expression of $IL-1{\beta}$, $TNF-{\alpha}$, MDA, $IL-1{\beta}$ mRNA, and $TNF-{\alpha}$ mRNA; (5) the infarction area of the hippocampus, and brain tissue injury in Alzheimer‘s diseased mice induced with ${\beta}A$ were investigated. Results : 1. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in BV2 microglia cell line treated with LPS. 2. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ protein production in BV2 microglia cell line treated with LPS. 3. For the NYDBT extract group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by $A{\beta}$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 4. The NYDBT extract suppressed the over-expression of $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, and CD68/CD11b, in the mice with Alzheimer's disease induced by $A{\beta}$. 5. The NYDBT extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by $A{\beta}$. 6. The NYDBT extract reduced the Tau protein, GFAP protein, and presenilin1/2 protein (immunohistochemistry) of hippocampus in the mice with Alzheimer's disease induced by $A{\beta}$. Conclusions : These results suggest that the NYDBT extract may be effective for the prevention and treatment of Alzheimer's disease.

• The Journal of Korean Medicine
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• v.43 no.2
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• pp.1-7
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• 2022

Objectives: Bee venom (BV) is a widely used therapy in Traditional East Asian Medicine (TEAM). We previously reported that BV was clinically effective for treating Parkinson's disease, that phospholipase A₂ (PLA₂) was the main component of BV, and that it induced regulatory T cells (Tregs) by binding CD206 on dendritic cells (DCs). Therefore, we aimed to reconfirm our findings in human blood samples and investigate the relationship between CD206⁺ DCs and clinical syndrome differentiation in TEAM. Methods: We surveyed 100 subjects with questionnaires on cold-heat patternization and obtained their blood samples. The obtained human peripheral blood monocytes (hPBMCs) were washed with phosphate-buffered saline (PBS). After resuspension with ex vivo media, numbers of cells were counted. Tregs were counted after culturing the samples in a 37℃ CO₂ incubator for 72 h. Results: We divided the subjects into a relatively high CD206⁺ group or a relatively low CD206⁺ group. The heat factor scores of high CD206⁺ group were significantly higher than that of low CD206⁺ group (high vs low: 239.2 ± 54.1 vs 208.4 ± 55.1, p=0.023). After culturing with PLA₂, Tregs increased in the high CD206⁺ group but decreased in the low CD206⁺ group. Conclusion: In this study, we reconfirm that CD206⁺ DCs induced Treg differentiation by incubating human blood samples with PLA₂ and that they showed an association with syndrome differentiation, especially with heat patterns, in TEAM. A heat pattern in TEAM might be one indication for PLA₂ therapy because its score was elevated in the high CD206⁺ group.

• The Korea Journal of Herbology
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• v.34 no.4
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• pp.9-18
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• 2019

Objectives : It has been known to be correlated between phlegm and dementia from the perspective of oriental medicine, but it is unexplored whether herbal medicines to treat phlegm have pharmacological actions on Alzheimer's disease (AD). The aim of this study was to evaluate and to compare effects of herbal medicines to treat phlegm against AD in vitro. Methods : We selected 11 herbal medicines which treat phlegm and obtained each extract by boiling in 10-fold distilled water for 2 h. And we performed the assay of acetylcholinesterase (AChE) inhibitory effects of 11 herbal extracts. Next, we evaluated neuroprotective effects of them against amyloid $beta_{25-35}$ ($A{\beta}_{25-35}$) plaque-induced toxicity in HT22 mouse hippocampal neuronal cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. To investigate whether they show the anti-inflammatory effects against lipopolysaccharide (LPS), we also measured the levels of nitric oxide (NO) in BV2 microglia cells using griess reagent assay. Results : We found that Gamiyeongsin-hwan (GYH) and Cheonghunhwadam-tang (CHT) exhibited remarkable AChE inhibitory effects. In HT22 cells, Arisaematis Rhizoma, Trichosanthis Semen and Fritillariae Thunbergii Bulbus suppressed $A{\beta}_{25-35}$ plaque-induced neuronal cell death. In BV2 cells, Cheongung-hwan significantly inhibited the increase of NO contents induced by LPS and GYH and CHT showed a tendency to inhibit LPS-induced NO generation. Conclusions : These results suggest that several herbal medicines to treat phlegm showed the significant effects on AChE inhibition, neuroprotection against $A{\beta}_{25-35}$ plaque-induced toxicity, and inhibition of NO generation. Therefore, we demonstrate the possibility that herbal medicines with treating phlegm has effects against AD.

• Korean Journal of Food Science and Technology
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• v.53 no.4
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• pp.423-433
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• 2021

This study was performed to examine the neuroprotective effect of the ethyl acetate fraction from Eucommia ulmoides oliver leaf (EFEL) on PM_2.5-induced cytotoxicity. EFEL had higher total phenolic and flavonoid contents than the other fractions. In ABTS and DPPH radical scavenging activities, the IC₅₀ of EFEL was measured as 212.80 and 359.13 ㎍/mL, respectively. To investigate the neuroprotective effect of EFEL, MTT and DCF-DA assays were performed on HT22, MC-IXC, and BV-2 cells. EFEL effectively decreased PM_2.5-induced intercellular reactive oxygen species (ROS) content and inhibited PM_2.5-induced cell death. In the results of protein expression related to cellular cytotoxicity on microglial cells (BV-2), EFEL had an improvement effect on cell apoptosis and inflammatory pathways. Rutin and chlorogenic acid were identified as the main physiological compounds. Moreover, it was expected that EFEL, including rutin and chlorogenic acid, could be functional food substances with neuroprotective effects against PM_2.5-induced oxidative stress.

• Journal of Physiology & Pathology in Korean Medicine
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• v.37 no.2
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• pp.36-44
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• 2023

Essential oils and aromatherapy have traditionally been used for the treatment of anxiety and depression with few side effects. This study aimed to investigate the effects of essential oils from six herbal prescriptions known to be effective in treating anxiety and depression in Korean medicine. The neuroprotective and anti-neuroinflammatory effects of six essential oils, including Gamisachil-tang (GMSCT), Guibi-tang (GBT), Sihogayonggolmoryeo-tang (SYM), Danchisoyosan (DCSYS), Sihosogansan (SHSGS), and Soyosan (SYS), were examined in PC12 and BV2 cells. In corticosterone (CORT)-stimulated PC12 cells, all six essential oils ameliorated the CORT-induced decrease in cell viability at a concentration of 10 ㎍/ml. GMSCT, GBT, and SHSGS recovered CORT-induced cytotoxicity at concentrations of 1 ㎍/ml and 10 ㎍/ml. In lipopolysaccharide (LPS)-stimulated BV2 cells, GBT (10 ㎍/ml) decreased interleukin (IL)-1β production, whereas SHSGS (1 ㎍/ml) inhibited tumor necrosis factor (TNF)-α production. In the MK-801-induced anxiety in zebrafish, electroencephalogram (EEG) assessment indicated that GMSCT and SHSGS induced recovery in the delta and beta power densities and reduced theta/beta and delta/beta ratios. DCSYS and SYS decreased theta power density and theta/beta ratio, whereas GBT and SYM showed no effects on EEG signals. In the tail suspension test (TST) in mice, GBT, DCSYS, SHSGS, and SYS exhibited antidepressant-like effects by decreasing immobility time. These results suggest that the essential oils from the six herbal prescriptions, except SYM, may have beneficial effects on anxiety and/or depression. Further studies should be conducted to investigate the molecular signaling pathways that mediate the effects of these essential oils on anxiety and depression.

• Journal of Oriental Neuropsychiatry
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• v.19 no.3
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• pp.69-84
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• 2008

Objective: This experiment was designed to investigate the effects of the KBHT hot water extract & ultra-fine powder on Alzheimer's Disease Model Induced by $\beta$A. Method: The effects of the KBHT hot water extract on expression of proinflammatory cytokine mRNA in BV2 microglial celll cell line treated by lipopolysacchaide(LPS). The effects of the KBHT hot water extract & ultra-fine powder on (1) the behavior (2) AChE in serum (3) the infarction area of the hippocampus, and brain tissue injury in Alzheimer's diseased mice induced with $\beta$A were investigated. Results: 1. The KBHT hot water extract suppressed the expression of proinflammatory cytokine mRNA in BV2 microglial cell line treated with LPS. 2. The KBHT hot water extract & ultra-fine powder a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by $\beta$A in the Morris water maze experiment, which measured stop-through latency and distance movement-through latency 3. The KBHT hot water extract & ultra-fine powder suppressed the over-expression of AChE activity in the serum of the mice with Alzheimer's disease induced by $\alpha$A. 4. The KBHT hot water extract & ultra-fine powder suppressed the expression of TNF-$\alpha$, IL-l$\beta$ protein significantly in the microglial cell of mice with Alzheimer's disease induced by 1$\beta$A. 5. The KBHT ultra-fine powder reduced infarction area of hippocampus significantly in the mice with Alzheimer's disease induced by $\beta$A. Conclusions: These results suggest that the KBHT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the PMCMT hot water extract & ultra-fine powder for Alzheimer's disease is suggested for future research.