• 생명과학회지
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• 제29권12호
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• pp.1371-1377
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• 2019

뇌에서 과량의 reactive oxygen species (ROS) 생성에 의해 유발되는 산화적 스트레스는 알츠하이머 질환과 같은 신경퇴행성 질환의 원인으로 알려져 있다. 본 연구에서는 kaempferol, kaempferol-3-O-glucoside, quercetin, quercetin-3-β-D-glucoside와 같은 flavonoid와 그 배당체의 H₂O₂ 유도 산화적 스트레스에 대한 C6 신경교세포보호 효과를 확인하였다. H₂O₂만을 처리한 control군은 아무것도 처리하지 않은 normal군에 비해 세포 생존율 감소와 ROS 생성 증가를 통해 C6 신경교세포의 산화적 손상이 유도되었음을 확인하였다. 반면 4가지 flavonoid를 각각 처리한 군의 경우, H₂O₂를 처리한 control군에 비해 세포 생존율 증가와 ROS 생성 감소를 통해 산화적 손상 억제를 통한 신경교세포 보호 효과를 확인하였다. Flavonoid의 신경교세포 보호 작용 메커니즘을 규명하기 위해, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and interleukine (IL)-1β 등의 염증 관련 단백질 발현을 측정하였다. H₂O₂를 처리한 control군은 normal군에 비해 iNOS, COX-2, IL-1β 단백질 발현 증가와 IκB-α 발현 감소를 통해 신경교세포의 산화적 손상으로 인한 염증 반응을 확인하였다. 반면, 4가지 flavonoid를 각각 처리한 군의 경우 iNOS, COX-2, IL-1β 단백질 발현 감소와 IκB-α 발현 증가를 나타내어, 염증 반응 개선을 통한 신경교세포 보호 효과를 확인하였다. 특히, quercetin과 그 배당체인 quercetin-3-β-D-glucoside를 처리한 군은 kaempferol과 그 배당체인 kaempferol-3-O-glucoside를 처리한 군에 비해 우수한 신경교세포 보호 효과를 나타내었다. 본 연구는 4가지 flavonoid가 신경교세포에서 산화적 스트레스 억제를 통해 신경퇴행성 질환을 예방 및 치료할 수 있을 것으로 사료된다.

• 한국식품영양학회지
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• 제25권4호
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• pp.800-806
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• 2012

체내에서 산화스트레스에 의해 생성되는 활성산소종(reacitve oxygen species, ROS)은 당뇨병, 비만 등과 같은 만성질환을 야기시키는 것으로 알려져 있다. 고려인삼(Panax ginseng)은 수천 년간 피로 회복 및 면역증강용 기능성식품으로 이용되어 왔고, 사포닌, 산성다당체, 페놀성 화합물과 같은 다양한 생리활성 물질을 함유하고 있다. 따라서 본 연구에는 고온, 고압처리하여 제조한 신규 인삼에 대한 산화적 스트레스 저감 효능을 평가하고자 하였다. C2C12 근육세포에 산화적 스트레스를 유도하기 위해 $H_2O_2$ 1 mM 처리하고, 전처리 조건을 달리한 인삼 시료를 처리하여 cell morphology 및 항산화 관련 유전자인 SOD, CAT 및 GPx를 살펴보았고, 3T3-L1 지방세포는 분화과정 중 ROS 생성 억제효과 및 CAT, GPx 및 Cu/Zn-SOD의 항산화효소 관련 유전자의 발현 정도를 조사하였다. 고온, 고압처리한 인삼은 산화적 스트레스가 유도된 C2C12 근육세포 및 3T3-L1 지방세포에서 유의적으로 산화적 스트레스를 저감하는 것으로 나타났다. 이상의 결과로 보아, 본 연구진에 의해 개발된 고온 및 고압 처리된 인삼은 항산화 및 항피로 효능이 기대되는 바이며, 본 연구는 동물세포 수준에서의 비교이며, 보다 정확한 작용기전의 구명을 위해 향후 추가적인 연구를 통한 비교 실험이 수행되어야 할 것으로 사료된다.

• Biomolecules & Therapeutics
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• 제23권2호
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• pp.156-164
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• 2015

Alzheimer's disease (AD) is a neurodegenerative disorder associated with progressive memory loss and neuronal cell death. Although numerous previous studies have been focused on disease progression or reverse pathological symptoms, therapeutic strategies for AD are limited. Alternatively, the identification of traditional herbal medicines or their active compounds has received much attention. The aims of the present study were to characterize the ameliorating effects of spinosin, a C-glucosylflavone isolated from Zizyphus jujuba var. spinosa, on memory impairment or the pathological changes induced through amyloid-${\beta}_{1-42}$ oligomer ($A{\beta}O$) in mice. Memory impairment was induced by intracerebroventricular injection of $A{\beta}O$ ($50{\mu}M$) and spinosin (5, 10, and 20 mg/kg) was administered for 7 days. In the behavioral tasks, the subchronic administration of spinosin (20 mg/kg, p.o.) significantly ameliorated $A{\beta}O$-induced cognitive impairment in the passive avoidance task or the Y-maze task. To identify the effects of spinosin on the pathological changes induced through $A{\beta}O$, immunohistochemistry and Western blot analyses were performed. Spinosin treatment also reduced the number of activated microglia and astrocytes observed after $A{\beta}O$ injection. In addition, spinosin rescued the $A{\beta}O$-induced decrease in choline acetyltransferase expression levels. These results suggest that spinosin ameliorated memory impairment induced through $A{\beta}O$, and these effects were regulated, in part, through neuroprotective activity via the anti-inflammatory effects of spinosin. Therefore, spinosin might be a useful agent against the amyloid ${\beta}$ protein-induced cognitive dysfunction observed in AD patients.

• The Korean Journal of Physiology and Pharmacology
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• 제17권2호
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• pp.175-180
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• 2013

Resveratrol is a phytoalexin found in grapes, red wine, and berries. Resveratrol has been known to have many beneficial health effects, such as anti-cancer, neuroprotective, anti-inflammatory, and life-prolonging effects. However, relatively little is known about the effects of resveratrol on the regulation of ligand-gated ion channels. We have previously reported that resveratrol regulates subsets of homomeric ligand-gated ion channels such as those of 5-$HT_{3A}$ receptors. The ${\gamma}$-aminobutyric $acid_C$($GABA_C$) receptor is mainly expressed in retinal bipolar cells and plays an important role in visual processing. In the present study, we examined the effects of resveratrol on the channel activity of homomeric $GABA_C$ receptor expressed in Xenopus oocytes injected with cRNA encoding human $GABA_C$ ${\rho}$ subunits. Our data show that the application of GABA elicits an inward peak current ($I_{GABA}$) in oocytes that express the $GABA_C$ receptor. Resveratrol treatment had no effect on oocytes injected with $H_2O$ or with $GABA_C$ receptor cRNA. Co-treatment with resveratrol and GABA inhibited $I_{GABA}$ in oocytes with $GABA_C$ receptors. The inhibition of $I_{GABA}$ by resveratrol was in a reversible and concentration-dependent manner. The $IC_{50}$ of resveratrol was $28.9{\pm}2.8{\mu}M$ in oocytes expressing $GABA_C$ receptor. The inhibition of $I_{GABA}$ by resveratrol was in voltage-independent and non-competitive manner. These results indicate that resveratrol might regulate $GABA_C$ receptor expression and that this regulation might be one of the pharmacological actions of resveratrol on the nervous system.

• 한국식품과학회지
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• 제46권4호
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• pp.483-488
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• 2014

브로콜리(Brassica oleracea var. italica) 잎의 n-hexane, chloroform, ethyl acetate, butanol 및 distilled water 분획물의 total phenolic 화합물 함량은 chloroform 분획물이 206.8 mg GAE/g으로 가장 높게 나타났으며, ABTS radical 소거활성 및 지질 과산화(MDA) 생성 억제효과 실험 결과에서 chloroform 분획물에서 가장 높은 활성을 나타내었다. in vitro 항산화 실험들에서 우수한 효과를 보인 chloroform 분획물을 이용하여 $H_2O_2$으로 유도한 산화적 스트레스에 대한 신경세포(PC12 cell)에서의 산화적 스트레스 생성 억제효과, 세포 생존율 그리고 세포막 손상 억제효과 역시 농도 의존적 경향을 나타내며 positive control 로 사용된 Vit.C와 유의적인 결과를 나타냈다. 마지막으로, HPLC 분석 결과 브로콜리 잎 chloroform 분획물에 존재하는 주요 phenolic 화합물은 feulic acid인 것으로 확인되었다. 본 연구 결과를 종합해볼 때, 생리활성 물질로서의 ferulic acid 등을 함유한 브로콜리 잎 chloroform 분획물은 in vitro 항산화 활성과 함께 신경세포 보호효과를 나타내는 고부가가치 기능성 소재로의 활용이 기대된다.

• Nutrition Research and Practice
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• 제12권3호
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• pp.199-207
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• 2018

BACKGROUND/OBJECTIVES: Fermented Laminaria japonica (FL), a type sea tangle used as a functional food ingredient, has been reported to possess cognitive improving properties that may aid in the treatment of common neurodegenerative disorders, such as dementia. MATERIALS/METHODS: We examined the effects of FL on scopolamine (Sco)- and ethanol (EtOH)-induced hippocampus-dependent memory impairment, using the Passive avoidance (PA) and Morris water maze (MWM) tests. To examine the underlying mechanisms associated with neuroprotective effects, we analyzed acetylcholine (ACh) and acetylcholinesterase (AChE) activity, brain tissue expression of muscarinic acetylcholine receptor (mAChR), cAMP response element binding protein (CREB) and extracellular signal-regulated kinases 1/2 (ERK1/2), and immunohistochemical analysis, in the hippocampus of mice, compared to current drug therapy intervention. Biochemical blood analysis was carried out to determine the effects of FL on alanine transaminase (ALT), aspartate transaminase (AST), and triglyceride (TG) and total cholesterol (TC) levels. 7 groups (n = 10) consisted of a control (CON), 3 Sco-induced dementia and 3 EtOH-induced dementia groups, with both dementia group types containing an untreated group (Sco and EtOH); a positive control, orally administered donepezil (Dpz) (4mg/kg) (Sco + Dpz and EtOH + Dpz); and an FL (50 mg/kg) treatment group (Sco + FL50 and EtOH + FL50), orally administered over the 4-week experimental period. RESULTS: FL50 significantly reduced EtOH-induced increase in AST and ALT levels. FL50 treatment reduced EtOH-impaired step-through latency time in the PA test, and Sco- and EtOH-induced dementia escape latency times in the MWM test. Moreover, anticholinergic effects of Sco and EtOH on the brain were reversed by FL50, through the attenuation of AChE activity and elevation of ACh concentration. FL50 elevated ERK1/2 protein expression and increased p-CREB (ser133) in hippocampus brain tissue, according to Western blot and immunohistochemistry analysis, respectively. CONCLUSION: Overall, these results suggest that FL may be considered an efficacious intervention for Sco- and EtOH-induced dementia, in terms of reversing cognitive impairment and neuroplastic dysfunction.

• 동의신경정신과학회지
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• 제22권4호
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• pp.201-218
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• 2011

Objectives : The aim of this study was to investigate the effects of Guibi-Tang(GBT) on focal brain ischemic injury induced by intraluminal filament insertion in rats. Methods : The ischemia was induced by intraluminal filament insertion into middle cerebral artery. Sprague-Dawley rats were divided into five groups, normal group(n=8); control group was ischemia induced and no treatment(n=8); GBT 1X group was ischemia induced and administrated 42.2 mg/ml/kg of Guibi-Tang orally(n=8); GBT 3X group was ischemia induced and administrated 126.6 mg/ml/kg of Guibi-Tang orally(n=8); GBT 6X group was ischemia induced and administrated 253.2 mg/ml/kg of Guibi-Tang orally(n=8) for 21 days. mGluR5, Bax, Bcl-2 and cytochrome C were investigated to observe the effects of Guibi-Tang on apoptosis. The effects of Guibi-Tang on neuroprotective/apoptotic agents in cresyl violet, choline acetyltranferase(ChAT) with ischemic injury were investigated. Results : The intensity of mGluR5 mRNA in the hippocampal CA1 was increased in normal and GBT(Guibi-Tang) 1X groups compared with the control group. The intensity of Bax mRNA in the hippocampal CA1 was decreased in normal and GBT 1X groups compared with the control group. However it was increased unexpectedly in GBT 3X group. The intensity of Bcl-2 mRNA in the hippocampal CA1 was increased in normal and GBT 1X groups compared with the control group. The intensity of Bax/Bcl-2 ratio in the hippocampal CA1 was decreased in normal and GBT 1X groups compared with the control group. The intensity of cytochrome C protein in the hippocampal CA1 was decreased in normal, GBT 1X and GBT 6X groups compared with the control group. The density of neurons stained by cresyl violet and ChAT was increased in normal and GBT 1X groups compared with the control group. Conclusions : These results suggest that Guibi-Tang may have protective effect on vascular dementia.

• Molecules and Cells
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• 제45권3호
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• pp.134-147
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• 2022

The anti-oxidant enzyme heme oxygenase-1 (HO-1) is known to exert anti-inflammatory effects. From a library of pyrazolo[3,4-d]pyrimidines, we identified a novel compound KKC080096 that upregulated HO-1 at the mRNA and protein levels in microglial BV-2 cells. KKC080096 exhibited anti-inflammatory effects via suppressing nitric oxide, interleukin1β (IL-1β), and iNOS production in lipopolysaccharide (LPS)-challenged cells. It inhibited the phosphorylation of IKK and MAP kinases (p38, JNK, ERK), which trigger inflammatory signaling, and whose activities are inhibited by HO-1. Further, KKC080096 upregulated anti-inflammatory marker (Arg1, YM1, CD206, IL-10, transforming growth factor-β [TGF-β]) expression. In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinetreated mice, KKC080096 lowered microglial activation, protected the nigral dopaminergic neurons, and nigral damage-associated motor deficits. Next, we elucidated the mechanisms by which KKC080096 upregulated HO-1. KKC080096 induced the phosphorylation of AMPK and its known upstream kinases LKB1 and CaMKKbeta, and pharmacological inhibition of AMPK activity reduced the effects of KKC080096 on HO-1 expression and LPS-induced NO generation, suggesting that KKC080096-induced HO-1 upregulation involves LKB1/AMPK and CaMKKbeta/AMPK pathway activation. Further, KKC080096 caused an increase in cellular Nrf2 level, bound to Keap1 (Nrf2 inhibitor protein) with high affinity, and blocked Keap1-Nrf2 interaction. This Nrf2 activation resulted in concurrent induction of HO-1 and other Nrf2-targeted antioxidant enzymes in BV-2 and in dopaminergic CATH.a cells. These results indicate that KKC080096 is a potential therapeutic for oxidative stress-and inflammation-related neurodegenerative disorders such as Parkinson's disease.

• Food Science and Biotechnology
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• 제17권2호
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• pp.279-286
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• 2008

The antioxidative activity of various enzymatic extracts from leaves of Perilla frutescens var. japonica was evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and alkyl radical scavenging activity using an electron spin resonance (ESR) spectrometer. For this study, the leaves were enzymatically hydrolyzed by 8 carbohydrases (Dextrozyme, AMG, Promozyme, Maltogenase, Termamyl, Viscozyme, Celluclast, and BAN) and 9 proteases [Flavourzyme, Neutrase, Protamex, Alcalase, PP-trypsin (trypsin from porcine pancreas), papain, pepsin, $\alpha$-chymotrypsin, and BP-trypsin (trypsin from bovine pancreas)]. The DPPH radical scavenging activities of Promozyme and Alcalase extracts were the highest, and the $IC_{50}$ values were 77.25 and $109.66\;{\mu}g/mL$, respectively. All enzymatic extracts of the leaves scavenged hydroxyl radical, and the $IC_{50}$ values of Celluclast and pepsin extracts which were the highest activity were 243.34 and $241.86\;{\mu}g/mL$, respectively. The BAN and $\alpha$-chymotrypsin extracts showed the highest scavenging activities, and the $IC_{50}$ values were 21.13 and $33.23\;{\mu}g/mL$, respectively. The pepsin extracts from the leaves showed protective effect on $H_2O_2$-induced DNA damage. In addition, the pepsin extracts decreased cell death in PC-12 cells against $H_2O_2$-induced oxidative damage. The findings of the present study suggest that enzymatic extracts of the leaves possess antioxidative activity.

• 동의생리병리학회지
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• 제16권1호
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• pp.141-145
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• 2002

In order to clarify the neuroprotective effect of Gamibojungikki-tang (GBJIKT) water extract on cultured mouse spinal sensory neuron damaged by glucose Oxidase (GO), MTT [3-(4,5-dimethylthiazole-2-yl) -2,5-diphenyltetrazolium bromide] assay and SRB (Sulforhodamine B) assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of GBJIKT water extract for 3 hours prior to exposure of GO. Cell viability of cultured mouse spinal sensory neurons exposed to various concentrations of GO for 8 hours was decreased in a dose-dependent manner. MTT50 values were 45 mU/ml GO. Cultured mouse spinal sensory neurons in the medium containing various concentration of GO for 8 hours showed decreasing of total protein synthesis. GO was toxic on cultured spinal sensory neurons. Pretreatment at GBJIKT water extract for 3 hours following GO prevented the GO-induced neurotoxicity such as decreasing of total protein synthesis. These results suggest that GO shows toxic effect on cultured spinal sensory neurons and GBJIKT water extract is highly effective in proecting the neurotoxicity induced by GO.