• Journal of Acupuncture Research
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• v.20 no.4
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• pp.85-92
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• 2003

목적 : 허혈시 발생되는 저산소중 상태에서는 세포독성을 유발한다고 알려져 있으나 정확한 기전은 아직 규명되지 않았다. 본 연구에서는 뇌허혈로 인한 세포독성의 기전을 유전자 발현을 통하여 살펴보고자 하였다. 방법 : 본 실험에서는 BV-2 microglia 세포주에 12시간 동안의 저산소 상태에서의 유전자 발현을 분석하기 위하여 마이크로에레이를 시행하였다. 결과 : 저산소 상태에서는 정상에 비하여 cathepsin F, growth factor independent 1, calcitonin/calcitonin-related poly, leucine-rich repeat LGI family membrane, dublecortin, cyclohydrolase 1, Ia-associated invariant chain, carbohydrate kinase-like과 erythrocyte protein band 4.1-like 3 등의 유전자 발현이 3배 이상 증가하였다. 한편 neuronal guanine nucleotide exchange factor, Bcl-2-related ovarian killer protein, chemokine (C-X-C motif) ligand 5, RNA binding motif protein 3, interleukin 2 receptor, alpha chain, crystallin zeta, cytochrome P450 subfamily IV B, asparagine synthetase과 moesin 등의 유전자 발현은 0.2배 이하로 감소하였다. 결론 : 이상의 결과는 저산소중에 관여하는 유전자 및 저산소중과 관련된 뇌경색 등의 질환의 기전을 밝히는데 기초적 자료로 이용될 수 있을 것이다.

• Korean journal of applied entomology
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• v.45 no.1 s.142
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• pp.31-36
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• 2006

Inflammation in the brain has known to be associated with the development of a various neurologiacal diseases. The hallmark of neuro-inflammation is the activation of microglia, brain macrophage. Pro-inflammatory compounds including nitric oxide(NO) are the main cause of neuro-degenerative disease such as Alzheimer's disease. In the study, we examined whether Harmonia axyridis extracts inhibit the NO production by a direct method using Griess reagent, western blotting and by RT-PCR(Reverse Transcription-Polymerase Chain Reactionin) the gene expression of inducible nitric oxide synthase(iNOS). Distilled water$(H_2O)$ and methanol(MeOH) extracts of H. axyridis inhibited the protein expression of TNF-a(Tumor Necrosis Factor) and IL-6(Interleukin) in LPS (Lipopolysaccharide) stimulated BV-2 cells at the concentration of 100 ng/ml. Incubation of BV-2 cells with the extracts of $H_2O$ of MeOH inhibited the LPS induced NO and iNOS protein. And this inhibition of iNOS protein is concordant with the inhibition of iNOS mRNA expression. These data suggested that H. axyridis extracts may play a crucial role in inhibiting the NO production.

• Journal of Physiology & Pathology in Korean Medicine
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• v.27 no.1
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• pp.92-98
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• 2013

Bee Venom(below BV) has been used in alternative medicine to treat the diseases, such as pain diseases. BV contains a variety of peptides, including melittin, apamin, adolapin, MCD peptide, enzymes(i.e. PLA2), amines(i.e. histamine and epinephrine), and nonpeptide components. The two main components of BV are melittin and PLA2. The cell cytotoxic effects through the activation of PLA2 by melittin have been suggested to be the critical mechanism for the depress of cancer cell. Melittin and PLA2 have been reported to induce apoptosis and to possess anti-cancer effects and neurite outgrowth in PC12 cells. Analysis of proliferation was confirmed by MTT assay. BV decreased cell number through dose- and duration-dependent manner and these effects are apparent at a concentration of 3 ${\mu}g/ml$. To observe which signaling molecules will be activated by BV, phosphorylation of ERK, p38 MAPK, JNK and ERM were examined by Western blot analysis. To study the long term effect of BV in human gastric adenocarcinoma cell lines, the image of cells treated with BV for 4 days were obtained. BV was shown to exhibit anti-cancer activity in human gastric adenocarcinoma cell lines at a broad range of concentrations of 3 ${\mu}g/ml$. ERK, p38 MAPK and JNK were found to increase in BV treated cells. However, ERM which known to be involved in the cell death, was gradually decreased to 30minutes after addition 3 ${\mu}g/ml$ of BV. These results provide a possible BV-induced inhibitory signal for cancer proliferation that is initiated by the decrease in ERM activity. Moreover, it is likely that the activation of ERK, p38 MAPK and JNK are required for the BV-induced inhibition of cancer proliferation.

• The Journal of Korean Medicine
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• v.29 no.4
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• pp.171-179
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• 2008

Objective: Bufonis Venenum is the traditional Korean medicine Chan Su, which is obtained from the skin and parotid venom gland of the toads. It has been used for myocardial diseases, inflammation diseases, pain relief, cancer and others. The main components of BV are cinobufotoxin, cinobufalin, bufalin and others. Of these, bufalin, the major active ingredient of BV, has been reported to induce apoptosis and to possess anti-tumor effects. There was no report of anti-tumor screening of BV on hepatic cancer and which signaling pathway can be involved. In order to examine the effect of BV on hepatic cancer and the related signaling pathway with BV-induced apoptosis, human Hep G2 cells were used. Methods: Analysis of apoptosis was confirmed by MTT assay. BV decreased cell viability in a dose and duration dependent manner. To observe which signaling molecules will be activated by BV, phosphorylation of MAPK (p38, ERK, JNK), caspase 8 and caspase 9 were examined by Western blot analysis. Results: The phosphorylation levels of p38 started to increase at 5 min after addition of 5 ${\mu}g$/ml of BV and sustained to increase until 48 hours. The phosphorylation levels of other MAPK (ERK and JNK), caspase 8 and caspase 9 increased in a time-dependent manner. These imply that BV may activate different signaling pathways, MAPK, caspase 8 and caspase 9. These results propose that BV may induce apoptosis on Hep G2 cells through the activation of MAPK, caspase 8 and caspase 9.

• Journal of Acupuncture Research
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• v.19 no.5
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• pp.136-148
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• 2002

본 연구는 osteoblastic cells에서 estogen 의 생합성을 유도하는 aromatase의 activity에 대한 봉독(蜂毒)작용을 측정하여, 봉독치료시 Arthritis의 진행 억제 및 estogen의 의한 bone formation의 효과여부를 검증하기 위해 실행하였다. 사용된 세포주로는 Osteoblastic phenotype으로 분화가 유도되는 Human leukaemic cell line FLG 29.1 및 the primary first-passage osteoblastic cells (hOB cells)이며, 이들을 각각 배양하고 각각의 RNA를 isolation한 뒤 PCR 증폭을 하였다. Aromatase에 대한 활성인자인 TPA와 TGF-${\beta}1$ 및 봉독을 이용하여 aromatase의 expression 및 activity에 대해 미치는 영향을 측정한 바, aromatase expression은 FLG 29.1 cell와 hoB cells에서, 50nM TPA 24시간 처리, 봉독 2 ~ 4시간 처리와 TGF-${\beta}1$ 3시간 처리로 유도한 결과 TPA와 TGF-${\beta}1$의 경우는 서로 유사하였고, 봉독에서 상대적으로 높게 나타났다. Aromatase activity는 FLG 29.1 cell, hoB cells에서 24시간 incubation한 결과, 모든 실험에서 일정하게 선상증가를 보였다. $5{\mu}{\ell}/m{\ell}$ BV에서 TPA와 TGF-${\beta}1$보다 뚜렷하게 증가하였으며, 0.5mM Bt2-cAMP, 50nM dexametasone처리에서는 유의성이 없었다. Estrogen 생합성을 촉매하는 aromatase activity BV가 처리에서 현저하게 증가하였기에, Rheumatis arthritis의 bone destruction에 대해 BV가 효과적인 역할을 할 것으로 보여진다.

• Korean Journal of Pharmacognosy
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• v.49 no.1
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• pp.1-6
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• 2018

Six compounds, eupatilin (1), dammaradienyl acetate (2), glutinol acetate (3), $3{\beta}-acetoxyoleanan-12-one$ (4), taraxasterol (5) and quercetin-3,4'-dimethyl ether (6) were isolated from the aerial parts of Artemisia absinthium. The chemical structures of compounds 1-6 were determined by the basis of physico-chemical properties and spectroscopic methods such as 1D and 2D NMR. Among them, compounds 2-5 were isolated from this plant for the first time. The inhibitory effects of these isolated compounds against nitric oxide (NO) production in LPS-induced RAW264.7 cells or BV2 microglia were also examined. Among the tested compounds, compound 1, eupatilin, inhibited the production of NO in LPS-induced RAW264.7 cells and BV2 microglia, respectively.

• The Journal of Internal Korean Medicine
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• v.30 no.4
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• pp.845-857
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• 2009

Background and Objectives : Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) have been reported to induce apoptosis in various cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. However, the comparative effect of VCA and BV on the anti-cancer effect and mechanisms of action has not been determined. In this study, the effect in a human hepatocellular carcinoma cell line, Hep G2 cells, was examined. Methods : Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in litro. The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action were examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. The involvement of kinase was examined in VCA or BV-induced apoptosis by using kinase inhibitors. Results : VCA and BV killed Hep G2 cells in a time and dose-dependent manner. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including MAPK/ERK, p38 MAPK and JNK. BV also activated PARP-1, MAPK/ERK. and p38 MAPK but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not by BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. VCA-induced apoptosis was partially inhibited by in the presence of JNK and p38 inhibitor, but BV only by p38 inhibitor. Conclusions : VCA-induced apoptosis is dependent on the activation of p38 and JNK. while BV-induced apoptosis is mediated by p38 activation in Hep G2 cells.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.20 no.2 s.33
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• pp.116-131
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• 2007

황기건중탕은 보비익기(補裨益氣) 및 거습열(祛濕熱)하는 효능(效能)으로 신체(身體)의 허약(虛弱)을 개선하며 염증(炎症)을 유발할 수 있는 외사(外邪)의 침입을 방어하여 인체의 면역기능(免疫機能)을 강화하는 처방으로 면역관련 세포중 하나인 microglial cell를 대상으로 면역세포에 황기건중탕이 어떠한 효능을 발휘하는 지를 살펴보았다. 물로 전탕(煎湯)한 황기건중탕을 여과한 후 실험한 결과 BV2 microglial cells에서 LPS로 자극하여 염증을 유발하는 주요 물질인 NO의 분비를 세포독성 없이 억제하였다. 또한NO를 생성하는 효소인 Cox-2의 발현도 감소시켰다. 그리고 기타 염증전구 물질들이 $TNF-{\alpha}$, $IL-1{\beta}$, IL-12도 황기건중탕의 용량이 증가함에 따라 의존적으로 감소함을 알 수 있었다. 이러한 결과를 보아 황기건중탕은 면역관련세포인 microglial cell에서 염증 관련 인자들의 분비 및 생성을 억제를 통하여 면역관련 상태를 개선시키는 것으로 사료된다.

• Korean Journal of Pharmacognosy
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• v.48 no.2
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• pp.108-112
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• 2017

To isolate and identify the constituents of the fruit galls of Actinidium polygama as well as evaluate the anti-inflammatory effects of the isolated compounds, a total of five triterpenes were isolated including $2{\alpha},3{\alpha},24$-trihydroxy-12-ursene-23-al-28-oic acid (1), corosolic acid (2), $2{\alpha},3{\alpha},24$-trihydroxyurs-12,20(30)-dien-28-oic acid (3), $2{\alpha},3{\alpha},24$-trihydroxyolean-12-en-28-oic acid (4), and $2{\alpha},3{\alpha},24$-trihydroxyurs-12-en-28-oic acid (pygenic acid B) (5). Compounds 1, 3, and 4 were isolated from the fruit galls of A. polygama for the first time. Compound 3 decreased NO production in LPS-stimulated RAW264.7 macrophages and mouse microglia BV2 cells in a concentration-dependent manner.

• Journal of Life Science
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• v.30 no.4
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• pp.331-342
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• 2020

The suppression of neuroinflammatory responses in microglial cells can be considered a key target for improving the progression of neurodegenerative diseases such as Alzheimer's disease (AD), Parkinson's disease (PD), and Huntington's disease (HD). Asparagus cochinchinensis has traditionally been used as a medicine to treat fever, cough, kidney disease, breast cancer, inflammatory diseases, and brain diseases. In this study, we investigated the neuroprotective mechanism of an aqueous extract from A. cochinchinensis root (AEAC), particularly its anti-inflammatory effects on lipopolysaccharide (LPS)-activated BV-2 microglial cells. BV-2 cells were treated with four different concentrations of AEAC. No significant toxicity was detected in BV-2 cells treated with AEAC. Nitric oxide (NO), cyclooxygenase-2 (COX-2) mRNA, and inducible nitric oxide synthase (iNOS) mRNA levels were 21% lower in the AEAC+LPS group than in the Vehicle+LPS group. Lower proinflammatory (TNF-α and IL-1β) and anti-inflammatory cytokine (IL-6 and IL-10) levels were also detected in the AEAC+LPS group than in the Vehicle+LPS group, albeit at varying rates. Moreover, the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group compared to the Vehicle+LPS group, enhancement of the phosphorylation of mitogen-activated protein kinase (MAPK) members after LPS treatment was significantly recovered in the AEAC-pretreated group, while cell cycle arrest at the G2/M phase caused by LPS treatment was less severe in the AEAC+LPS group. The increase in reactive oxygen species (ROS) generation induced by LPS treatment was also lower in the AEAC-pretreated group than in the Vehicle+LPS group. This is the first study to show that AEAC exerts anti-neuroinflammatory activity against LPS stimulation by regulating the MAPK signaling pathway, the cell cycle, and ROS production.