• 대한본초학회지
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• 제34권5호
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• pp.29-37
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• 2019

Objectives : Bang-poong (Saposhnikovia divaricata; SD) was traditionally used to treat inflammatory disorders. In this study, we aimed to investigate whether Bang-poong and related species including SD, Glehnia littoralis (GL), and Peucedanum japonicum (PJ) possess neuroprotective effects and acetylcholinesterase (AChE) inhibitory activities. Methods : Roots of SD, GL and PJ were extracted with distilled water (DW) or 70% ethanol (EtOH). We assessed 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities of the extracts. To examine neuroprotective effects, we measured cell viability in PC12 or HT22 cells after treatment of the extracts with $H_2O_2$ or amyloid-beta ($A{\beta}$). To assess anti-neuroinflammatory effects, we measured the nitric oxide (NO) levels after treatment with the extracts and lipopolysaccharide (LPS) in BV2 microglial cells. In addition, we performed AChE inhibition assay to explore effects of the extracts on the cholinergic system. Results : DW and EtOH extracts of SD, GL and PJ showed mild DPPH free radical scavenging activities. Also, DW extracts of GL and PJ showed protective effects against $H_2O_2$-induced toxicity in PC12 cells. In LPS-activated BV2 cells, EtOH extracts of SD, GL and PJ exerted inhibitory effects on NO production. Meanwhile, DW extracts of SD, GL and PJ inhibited the $A{\beta}$-induced cell death in HT22 cells. In addition, DW and EtOH extracts of GL exhibited remarkable inhibitory activities on AChE. Conclusions : We demonstrated that SD, GL and PJ exert anti-oxidative, anti-neuroinflammatory and AChE inhibitory activities. These results indicate that SD, GL and PJ could be potential candidates for neurological disorders.

• 대한본초학회지
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• 제39권5호
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• pp.31-38
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• 2024

Objectives : Achyranthes japonica Nakai (AJ) and Achyranthes bidentata Blume (AB) have been used without distinguishment. Moreover, comparative studies of AJ and AB on physiological activity in the organism levels remain fully understood. In this study, we aimed to evaluate and compare the effects of AJ and AB on anti-neuroinflammatory and neuroprotective effects. Methods : AJ and AB were extracted with distilled water (DW) and 70% ethanol (EtOH) extract. For the evaluation of anti-neuroinflammatory effects, we measured the production of nitric oxide (NO) in lipopolysaccharide (LPS)-treated BV2 microglial cells. To evaluate the neuroprotective effects, we assessed cell viability against toxicity, including hydrogen peroxide (H₂O₂), 6-hydroxydopamine (6-OHDA), and amyloid-beta (A𝛽), respectively, in PC12 or HT22 cells. Results : DW and 70% EtOH extracts of AJ and AB inhibited LPS-induced NO production in BV2 cells, with no significant differences between the origins and extraction solvents. Additionally, AJ and AB had no cytotoxicity, and exhibited the similar neuroprotective effects against H₂O₂ and 6-OHDA toxicities in PC12 cells, showing stronger activity in 70% EtOH extract compared to the DW extract. Furthermore, 70% EtOH extracts of AJ and AB protected neuronal cell against A𝛽 toxicity-induced cytotoxicity in HT22 cells. Conclusions : We demonstrated that AJ and AB have anti-neuroinflammatory and neuroprotective effects in the 70% EtOH extract compared to DW extract, with no significant differences between the species. These results suggested that AJ and AB would be the potential candidates for neurodegenerative diseases.

• Mass Spectrometry Letters
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• 제5권3호
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• pp.70-78
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• 2014

Microglia are the confined immune cells of the central nervous system (CNS). In response to injury or infection, microglia readily become activated and release proinflammatory mediators that are believed to contribute to microglia-mediated neurodegeneration. In the present study, inflammation was induced in the immortalized murine microglial cell line BV-2 by lipopolysaccharide (LPS) treatment. We firstly performed phosphoproteomics analysis and phosphoinositide lipidomics analysis with LPS activated microglia in order to compare phosphorylation patterns in active and inactive microglia and to detect the pattern of changes in phosphoinositide regulation upon activation of microglia. Mass spectrometry analysis of the phosphoproteome of the LPS treatment group compared to that of the untreated control group revealed a notable increase in the diversity of cellular phosphorylation upon LPS treatment. Additionally, a lipidomics analysis detected significant increases in the amounts of phosphoinositide species in the LPS treatment. This investigation could provide an insight for understanding molecular mechanisms underlying microglia-mediated neurodegenerative diseases.

• 한국식품영양학회:학술대회논문집
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• 한국식품영양학회 2003년도 하계 학술 심포지엄
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• pp.82-82
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• 2003

오디는 옛부터 보혈자음(補血滋陰), 생진윤조(生津閏操), 현훈이명(眩最耳鳴), 심계실면(心悸失眠), 수발조백(鬚髮早白) 등을 치료하는 효능을 가진다고 알려져 있다. 상심자(桑 子)라 하여 뽕나무 열매로서 한방에서 상심(桑 ), 상실(桑實), 오심(烏 ), 흑심(黑 ) 등으로 지칭되며, 뽕나무과(Moraceae)에 속하는 뽕나무(Morus alba L)의 성숙한 과실로 취화과(聚花果)에 속하며 작은 수과(瘦果)가 많이 모여 이루어진 장원형으로 길이 1 내지 2 cm, 지름 0.5 내지 0.8 cm이며 황갈색, 갈홍색 또는 암자색을 띠고 짧은 줄기가 있다. 작은 수과(瘦果)는 난원형으로 조금 납작한 편이며, 길이는 약 2 mm, 너비는 약 1 mm 이고 육질의 화편(花片) 4개가 둘러싸고 있다. 그러나 아직까지는 오디에 대하여 자연과학적인 연구방법을 이용하여 구체적으로 연구된 바가 많지 않다. 또한 이의 생리활성 성분에 대해서도 그 작용과 연관지어 보고된 바가 많지 않다. 뇌졸중 가운데 뇌조직으로 가는 혈액 공급의 감소 혹은 차단으로 발생되는 허혈성 뇌졸중(ischemic stroke)은 전체 뇌졸중 환자의 약 80%정도를 차지하고 있으나 아직까지 뇌신경세포 손상기전의 복잡성 등으로 뇌졸중으로 발생하는 뇌신경세포의 손상을 보호해 줄 수 있는 물질이 개발되어 있지 못한 실정이다. 한편, 천연물로부터 뇌허혈 보호작용을 가지는 물질의 도출은 주로 한방에서 처방을 중심으로 이루어지고 있으며, 따라서 처방으로부터 신경보호작용을 가지는 물질의 도출은 그 처방에 함유되어 있는 각종 생약이 갖는 다양한 활성으로 인해 어려운 점이 있으며, 비록 효과가 있다고 하더라도 과학적인 입증자료가 매우 미비한 실정이다. 이에 본 연구에서는 위에 언급된 자료를 토대로 오디추출물이 뇌허혈에 효과가 있을 것이라는 가정하에 in vitro system을 이용하여 오디추출물의 신경보호작용기전을 검색하고자 DPPH radical 의 생성억제효과, PC12 cell line을 이용한 산화적 스트레스에 의한 오디추출물의 방어효과, LSP에 의하여 활성화된 BV-2 cell에 미치는 영향 등을 검색하였다. 오디의 DPPH 소거활성은 단일화랍물인 bacicalein과 유사한 효과를 나타냈으며 PC12 cell line을 이용한 산화적 스트레스에 의한 오디의 방어 효과는 LDH activity를 행한 결과 농도 의존적으로 LDH 유리 량을 감소시켰다. 뇌손상시에 오디추출물의 염증방어 효과를 관찰하기 위하여 microglial cell line인 BV2 세포주를 선택하여 배양한 후 LPS로 자극을 준 후 일차적인 염증지표인 NO양을 측정한 결과 BV2 cell에 LPS 100 ng/ml을 처리하는 경우 nitrite량이 유의적으로 증가하였는데 이때 오디추출물을 1, 5, 10 ug/ml의 용량으로 처리하는 경우 역시 저농도에서 NO생성량을 감소시켰다. 정리해보면 뽕나무과 식물인 오디는 신경보호효과가 있는 것으로 사료되는데 이러한 효과는 아마도 오디의 항산화효과에 기인한 것으로 추정된다.

• Journal of Ginseng Research
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• 제41권2호
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• pp.151-158
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• 2017

Background: Chong-Myung-Tang (CMT) extract is widely used in Korea as a traditional herbal tonic for increasing memory capacity in high-school students and also for numerous body ailments since centuries. The use of CMT to improve the learning capacity has been attributed to various plant constituents, especially black ginseng, in it. Therefore, in this study, we have first investigated whether black ginseng-enriched CMT extracts affected spatial learning using the Morris water maze (MWM) test. Their molecular mechanism of action underlying improvement of learning and memory was examined in vitro. Methods: We used two types of black ginseng-enriched CMT extracts, designated as CM-1 and CM-2, and evaluated their efficacy in the MWM test for spatial learning behavior and their anti-inflammatory effects in BV2 microglial cells. Results: Our results show that both black ginseng-enriched CMT extracts improved the learning behavior in scopolamine-induced impairment in the water maze test. Moreover, these extracts also inhibited nitric oxide production in BV2 cells, with significant suppression of expression of proinflammatory cytokines, especially inducible nitric oxide synthase, cyclooxygenase-2, and $interleukin-1{\beta}$. The protein expression of mitogen-activated protein kinase and nuclear $factor-{\kappa}B$ pathway factors was also diminished by black ginseng-enriched CMT extracts, indicating that it not only improves the memory impairment, but also acts a potent anti-inflammatory agent for neuroinflammatory diseases. Conclusion: Our research for the first time provides the scientific evidence that consumption of black ginseng-enriched CMT extract as a brain tonic improves memory impairment. Thus, our study results can be taken as a reference for future neurobehavioral studies.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2018년도 춘계학술발표회
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• pp.89-89
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• 2018

Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In this study, anti-inflammatory effects of Abeliophyllum distichum stem (ADS) ethyl acetate extract were examined. Furthermore, possible molecular mechanisms of the anti-inflammatory effects were dissected. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 were investigated by Western blot analysis. The extract of ADS significantly decreased the production of pro-inflammatory cytokines. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. Our findings provide evidence for the popular use of Abeliophylli distichum in inflammation around Goesan region and also suggest that the stem extract has potential therapeutic benefits against several inflammatory diseases.

• Biomolecules & Therapeutics
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• 제22권5호
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• pp.414-419
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• 2014

Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that regulate cell-matrix composition and are also involved in processing various bioactive molecules such as cell-surface receptors, chemokines, and cytokines. Our group recently reported that MMP-3, -8, and -9 are upregulated during microglial activation and play a role as proinflammatory mediators (Lee et al., 2010, 2014). In particular, we demonstrated that MMP-8 has tumor necrosis factor alpha (TNF-${\alpha}$)-converting enzyme (TACE) activity by cleaving the prodomain of TNF-${\alpha}$ and that inhibition of MMP-8 inhibits TACE activity. The present study was undertaken to compare the effect of MMP-8 inhibitor (M8I) with those of inhibitors of other MMPs, such as MMP-3 (NNGH) or MMP-9 (M9I), in their regulation of TNF-${\alpha}$ activity. We found that the MMP inhibitors suppressed TNF-${\alpha}$ secretion from lipopolysaccharide (LPS)-stimulated BV2 microglial cells in an order of efficacy: M8I>NNGH>M9I. In addition, MMP inhibitors suppressed the activity of recombinant TACE protein in the same efficacy order as that of TNF-${\alpha}$ inhibition (M8I>NNGH>M9I), proving a direct correlation between TACE activity and TNF-${\alpha}$ secretion. A subsequent pro-TNF-${\alpha}$ cleavage assay revealed that both MMP-3 and MMP-9 cleave a prodomain of TNF-${\alpha}$, suggesting that MMP-3 and MMP-9 also have TACE activity. However, the number and position of cleavage sites varied between MMP-3, -8, and -9. Collectively, the concurrent inhibition of MMP and TACE by NNGH, M8I, or M9I may contribute to their strong anti-inflammatory and neuroprotective effects.

• 생명과학회지
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• 제27권1호
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• pp.8-14
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• 2017

치자 열매는 아시아에서 음식과 옷의 염료로 사용되어 왔다. 본 연구에서는 BV-2 소교세포에서 치자열매 열수추출물(GJ)의 항염증 효과를 관찰하고 그 작용 기전을 연구하였다. GJ는 세포에 독성을 유도하지 않으면서 lipopolysaccharide로 인한 nitric oxide (NO) 분비와 inducible nitric oxide synthase (iNOS) 생성 및 활성산소 생성을 억제하였다. 또한 GJ는 농도의존적으로 heme oxygenase-1 (HO-1)의 발현을 유도하였다. 더군다나 HO-1 siRNA를 처리했을 때는 GJ가 iNOS의 발현을 억제하지 못하였다. GJ는 HO-1의 발현에 관여하는 전사인자인 nuclear factor E2-related factor 2를 핵으로 이동시켰다. 또한 GJ에 의한 HO-1의 발현은 phosphatidylinositol 3-kinase(PI-3K) 및 p38 kinase 억제제에 의해 감소되었으며, GJ가 Akt와 p38 kinase의 인산화를 유도하였다. 이상의 결과를 종합해보면, GJ는 PI3K/Akt 및 p38 신호전달과정을 통해 HO-1의 발현을 유도함으로써 NO와 같은 염증매개물질의 생성을 억제한다는 것을 알 수 있다. 이러한 연구결과는 치자열매가 신경염증을 억제하는 새로운 기전을 밝힌 것이다.

• Journal of Nutrition and Health
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• 제50권1호
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• pp.25-31
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• 2017

퇴행성 뇌신경 질환의 원인이 되는 것으로 알려진 미세아교세포의 과도한 활성화에 의한 신경염증반응에 미치는 미역쇠의 보호 효과를 알아보기 위해 LPS를 처리한 BV2 세포에서 미역쇠에서 얻은 에탄올 추출물을 이용하여 실험을 수행하였다. 미세아교세포의 활성화를 유도하는 LPS의 처리는 신경염증반응의 지표인 NO의 생성량과 이들을 조절하는 iNOS, COX-2의 발현을 증가시켰다. 미역쇠 추출물의 처리는 LPS가 유도하는 NO의 생성량을 농도 의존적으로 억제하였고 iNOS와 COX-2의 발현을 억제하여 NO 생성량 저해와 유사한 양상의 결과를 나타내었다. 미역쇠 추출물의 신경 염증반응 저해 효과가 $NF-{\kappa}B$의 활성화 조절을 통해 일어나는지를 알아보기 위해 $NF-{\kappa}B$의 핵으로의 전이, $I{\kappa}B$의 인산화, $NF-{\kappa}B$ 억제제인 PDTC를 이용한 NO의 생성량에 미치는 효과를 확인하였다. 미역쇠 추출물 처리에 의해 핵분획물에서의 $NF-{\kappa}B$ 발현은 현저히 감소하였고 $I{\kappa}B$의 인산화를 억제하였으며 PDTC의 처리로 NO의 생성량은 감소하였다. 이상의 결과는 미세아교세포의 활성화로 인해 발생되는 신경염증반응에 미역쇠 추출물이 $NF-{\kappa}B$의 활성 억제를 통해 NO의 생성을 저해함으로써 항신경염증 효과가 있음을 보여주는 것으로 미역쇠 추출물이 신경염증 관련 뇌신경 질환의 제어하는데 있어서 치료효과를 가지는 소재로서 이용 가능성에 대한 정보를 제공할 것으로 사료된다.

• 동의생리병리학회지
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• 제22권6호
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• pp.1572-1578
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• 2008

In previous report, Seungmagalgeun-tang (SGT) could exert its anti-inflammatory actions in the BV-2 microglial cells. However, study on the anti-inflammatory effect of SGT in mast cells has not been identified. Therefore, we examined on the anti-inflammatory effect of SGT on the phorbol 12-myristate 13-acetate (PMA) and calcium ionophore A23187-induced rat basophilic leukemia (RBL-2H3) cells. SGT inhibited the release of ${\beta}$-hexosaminidase and secretion and expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-4 on RBL-2H3 cells, without affecting cell viability. The protein expression level of nuclear factor (NF)-${\kappa}B$ (p65) was decreased in the nucleus by SGT. In addition, SGT suppressed the degradation of inhibitory protein $I{\kappa}B-{\alpha}$ protein, the activation of p38 mitogen-activated protein kinase (MAPK), and the expressions of cyclooxygenase (COX)-2 mRNA and protein level in RBL-2H3 cells. These results suggest that SGT could be involved anti-allergic effect by control of NF-${\kappa}B$ (p65) translocation into the nucleus through inhibition of $I{\kappa}B-{\alpha}$ degradation and suppression of COX-2 expression.