• 한국식품영양과학회지
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• 제45권7호
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• pp.958-965
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• 2016

본 연구는 산업 부산물의 이용성 증진을 위하여 폐자원인 녹차씨껍질 추출물의 항산화 활성 및 신경세포 보호 효과에 관하여 평가하였다. 녹차씨껍질로부터 유용성 성분을 얻기 위하여 에탄올 추출을 한 결과 약 1.44%의 추출물의 수율을 얻을 수 있었고, 항산화 활성에 관하여 평가하기 위하여 라디칼 소거능 및 xanthine oxidase 저해능, 환원력을 평가한 결과, 녹차씨껍질 추출물의 농도가 증가할수록 항산화 활성이 유의적으로 증가하는 것으로 나타났다. 또한, 이러한 녹차씨껍질 추출물의 뇌신경세포 보호 효과에 관하여 알아보기 위하여 생쥐의 해마 유래 뇌신경세포에 녹차씨껍질 추출물을 처리한 후 $H_2O_2$로 산화적인 스트레스를 유도하여 세포독성에 관하여 알아본 결과, 녹차씨껍질 추출물의 처리는 농도 의존적으로 뇌신경 세포의 생존율을 증가시켰으며, 이에 따라 항산화 효소인 SOD 활성이 증가하고 지질과산화 생성물인 MDA level이 감소한 것을 알 수 있었다. 이상의 결과들로 녹차씨껍질 추출물의 항산화 활성 및 뇌신경세포 보호 효과에 관하여 확인할 수 있었으며, 추후 어떤 메커니즘으로 신경세포를 보호하는지 추가적인 연구가 필요할 것으로 생각한다. 또한, 이러한 결과를 활용하여 녹차씨껍질을 기능성 소재로 활용한다면 폐자원인 녹차씨껍질을 재활용하는 차원에서 그 경제적 가치가 매우 클 것으로 생각한다.

• 대한본초학회지
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• 제34권4호
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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.

• 생약학회지
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• 제45권3호
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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.

• 한국식품위생안전성학회지
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• 제38권6호
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• pp.430-441
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• 2023

녹용은 수많은 연구에서 면역력 강화를 포함한 영양 및 의학적 가치를 입증하였으며 전통적인 약으로 널리 사용되고 있다. 본 연구는 녹용 추출물 (sample 1: 생녹용 추출물, sample 2: 건녹용 추출물, sample 3: 동결 건조 추추물)의 일반성분과 우론산, 황산화 글리코사미노글리칸, 시알릭산, 콜라겐을 포함한 유효성분을 조사하고, 액체 크로마토그래피quadrupole-time-of-light mass spectrometry (UPLC/QTOFMS)를 사용하여 녹용 추출물의 화학 성분을 분석하는데 목적이 있다. 또한, HT22 해마 세포, BV2 미세아교세포, RAW264.7 대식세포 및 HaCaT 케라틴 세포를 사용하여 MTT 분석을 통해 녹용 추출물의 세포 독성 효과 평가와 암컷과 수컷 ICR 마우스에 녹용 추출물을 각각 (0, 500, 1000, 2000 mg/kg) 경구투여 하여 급성 독성평가를 실시하였다. 투여후에는 OECD 가이드라인에 따라 마우스의 일반독성, 생존율, 체중 변화, 사망률, 임상 징후 및 부검 결과를 관찰하였다. 결과적으로 녹용 추출물은 HaCaT 케라틴 세포에서 세포 독성 효과가 없었으며, 건녹용 추출물에서는 HT22 해마 세포에서 500 ㎍/mL, RAW264.7 대식세포의 경우 1000 ㎍/mL 에서, 동결건조추출물에서는 RAW264.7 세포와 BV2 미세아교세포의 경우 500 ㎍/mL 및 1000 ㎍/mL 농도에서 세포 독성을 가지고 있음을 보였다. 그러나 마우스를 이용한 급성 독성 평가에서는 녹용 추출물 시료를 처리한 모든 마우스에서 사망률, 임상 징후 및 부검 결과 특이사항이 없었으며 이는 LD50이 2000 mg/kg 이상으로 사료된다. 그러나 인간에 대한 안전성에 대한 충분한 증거를 확보하기 위해서는 동물과 사람에 대한 추가적인 연구가 필요하다.

• BMB Reports
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• 제56권4호
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• pp.234-239
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• 2023

Thioredoxin-like protein 1 (TXNL1), one of the thioredoxin superfamily known as redox-regulator, plays an essential in maintaining cell survival via various antioxidant and anti-apoptotic mechanisms. It is well known that relationship between ischemia and oxidative stress, however, the role of TXNL1 protein in ischemic damage has not been fully investigated. In the present study, we aimed to determine the protective role of TXNL1 against on ischemic injury in vitro and in vivo using cell permeable Tat-TXNL1 fusion protein. Transduced Tat-TXNL1 inhibited ROS production and cell death in H₂O₂-exposed hippocampal neuronal (HT-22) cells and modulated MAPKs and Akt activation, and pro-apoptotic protein expression levels in the cells. In an ischemia animal model, Tat-TXNL1 markedly decreased hippocampal neuronal cell death and the activation of astrocytes and microglia. These findings indicate that cell permeable Tat-TXNL1 protects against oxidative stress in vitro and in vivo ischemic animal model. Therefore, we suggest Tat-TXNL1 can be a potential therapeutic protein for ischemic injury.

• 대한한의학회지
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• 제39권3호
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• pp.1-16
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• 2018

Objectives: The purpose of this study was to investigate the effects of 56 herbal formulae covered by the National Health Insurance Corporation (NHIC) on dementia-related biomarkers and neuronal cell changes. Methods: The 56 herbal formulae were extracted with 70% ethanol at $100^{\circ}C$ for 2 h. The antioxidant properties was measured by radical scavenging assay using ABTS+ radical. The acetylcholinesterase (AChE) activity was tested by Ellman's assay and $amyloid-{\beta}$ ($A{\beta}$) aggregation was determined using fluorescence method. To estimate the inhibitory effects of herbal formulae on neuronal cell death and inflammation using HT22 hippocampal cells and BV-2 microglia, respectively. Results: Among the 56 herbal formulae, Dangguiyukhwangtang, Banhasasimtang, Samhwangsasimtang, Cheongwiesan, Hwangryunhaedoktang, Banhabaekchulchunmatang, Jaeumganghwatang, Cheongseoikgitang, and Hoechunyanggyuksan has a significant inhibitory effects on acetylcholinesterase (AChE) activity. Doinseunggitang and Samhwangsasimtang exerted the effect on the inhibition of $amyloid-{\beta}$ ($A{\beta}$) aggregation. Additionally, 10 herbal formulae affected AChE and $A{\beta}$ aggregation revealed antioxidant activity as well as neuroprotective and anti-neuroinflammation effects in neuronal cell lines. Conclusions: 10 herbal formulae that have been shown to be effective against the major dementia markers have been shown to have antioxidant activity, neuronal cell protection and inhibition of brain inflammation. Further investigation of these herbal formulae will need to be validated in dementia animal models.

• Journal of Microbiology and Biotechnology
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• 제28권1호
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• pp.12-21
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• 2018

Photorhabdus temperata (PT), a gram-negative bacterium, lives symbiotically within entomopathogenic nematodes. The insecticidal compounds derived from Photorhabdus are used as biopesticides in agriculture. However, the physiological properties are not well characterized. In the course of our screening for neuroprotective and anti-neuroinflammatory substances from natural products, the culture broth of PT showed considerable activities. By activity-guided purification, five anthraquinones, namely, 3-methoxychrysazine (1), 1,3-dimethoxy-8-hydroxy-9,10-anthraquinone (2), 1,3,8-trihydroxy-9,10-anthraquinone (3), 3,8-dihydroxy-1-methoxy-9,10-anthraquinone (4), and 1,3,4-trimethoxy-8-hydroxy-9,10-anthraquinone (5), were isolated from the ethyl acetate fraction of the PT culture broth. Among the isolated compounds, $75{\mu}M$ 3 significantly protected mouse hippocampal neuronal cells (HT22) against 5 mM glutamate-induced cell death via the inhibition of reactive oxygen species production, $Ca^{2+}$ influx, and lipid peroxidation. Additionally, 3 and 4 effectively suppressed the interferon-${\gamma}$-induced neuroinflammation of mouse-derived microglial cells (BV2) at 10 ng/ml, via the reduction of nitric oxide, interleukin-6, and tumor necrosis factor-${\alpha}$. Anthraquinones 3 and 4 derived from the PT culture broth are a potential starting point to discover neuroprotective and anti-neuroinflammatory drug leads. The novel compound 5 is reported for the first time in this study.

• Preventive Nutrition and Food Science
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• 제21권3호
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• pp.221-226
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• 2016

A new type of doenjang was manufactured by mixing soaked soybean, koji (Rhizopus oryzae), cheonggukjang (Bacillus amyloliquefaciens MJ1-4 and B. amyloliquefaciens EMD17), and Pichia farinosa SY80 as a yeast, salt, and water, followed by fermentation with koji that was made by fermenting whole wheat with R. oryzae. The mixed culture doenjang was designed to have a more palatable flavor and stronger biological activities than the conventional product. The extract of mixed culture doenjang showed higher antioxidant activity than the commercial doenjang as evaluated by the ferric reducing antioxidant power assay although it was not significantly different from the commercial product in 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activities. Further, the mixed culture doenjang reduced intracellular reactive oxygen species levels and protected cells from glutamate-induced cytotoxicity more efficiently in human hippocampal HT22 neuroblastoma cells than the commercial doenjang. In conclusion, a newly-developed mixed culture doenjang had a strong antioxidant activity in vitro and cultured cell model systems, exhibited a potential to prevent oxidative stress-associated disorders although animal and clinical studies are needed to confirm its in vivo efficacy.

• 약학회지
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• 제55권2호
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• pp.121-126
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• 2011

Sipjundaebo-tang was a well-known restorative traditional herbal prescription that used to treat anemia, anorexia, fatigue and inflammation. In this study, we examined the bioconversion of compounds in the Sipjundaebo-tang (SJ) and fermented Sipjundaebo-tang with Lactobacillus fermentum KFRI 164 (FSJ) using established HPLC-DAD method. The chromatogram of FSJ has shown that the contents of six bioactive compounds 5-HMF, paeoniflorin, ferulic acid, cinnam aldehyde, decursin, glycyrrhizin in SJ has decreased. And the contents of unknown compounds (1), (2), (3), (4) and (5) in FSJ were higher than each contents of SJ. The antioxidant activities of SJ and FSJ were conducted by DPPH free radical and Hydrogen peroxide ($H_2O_2$) scavenging assay. Electron donating activity (EDA, %) value of FSJ has shown higher than 21.9% and 14.5% at a concentration of 0.5 mg/ml for DPPH radical scavenging activity and $H_2O_2$ scavenging activity, respectively. Also, the neuroprotective activities of SJ and FSJ against glutamate-induced oxidative stress in a mouse hippocampal cell line (HT22) were evaluated by MTT assay. As a result, FSJ has shown higher neuroprotective activity than 56.5% comparing with SJ. In conclusion, the fermented SJ using microorganism could convert compounds in SJ and enhance antioxidant activity and neuroprotective activity.

• Journal of Ginseng Research
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• 제44권3호
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• pp.442-452
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• 2020

Backgroud: Sleep deprivation (SD) impairs learning and memory by inhibiting hippocampal functioning at molecular and cellular levels. Abnormal autophagy and apoptosis are closely associated with neurodegeneration in the central nervous system. This study is aimed to explore the alleviative effect and the underlying molecular mechanism of stem-leaf saponins of Panax notoginseng (SLSP) on the abnormal neuronal autophagy and apoptosis in hippocampus of mice with impaired learning and memory induced by SD. Methods: Mouse spatial learning and memory were assessed by Morris water maze test. Neuronal morphological changes were observed by Nissl staining. Autophagosome formation was examined by transmission electron microscopy, immunofluorescent staining, acridine orange staining, and transient transfection of the tf-LC3 plasmid. Apoptotic event was analyzed by flow cytometry after PI/annexin V staining. The expression or activation of autophagy and apoptosis-related proteins were detected by Western blotting assay. Results: SLSP was shown to improve the spatial learning and memory of mice after SD for 48 h, accomanied with restrained excessive autophage and apoptosis, whereas enhanced activation of phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin signaling pathway in hippocampal neurons. Meanwhile, it improved the aberrant autophagy and apoptosis induced by rapamycin and re-activated phosphoinositide 3-kinase/Akt/mammalian target of rapamycin signaling transduction in HT-22 cells, a hippocampal neuronal cell line. Conclusion: SLSP could alleviate cognitive impairment induced by SD, which was achieved probably through suppressing the abnormal autophagy and apoptosis of hippocampal neurons. The findings may contribute to the clinical application of SLSP in the prevention or therapy of neurological disorders associated with SD.