• 생명과학회지
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• 제24권1호
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• pp.86-91
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• 2014

Trichoplusia ni (T. ni) 세포는 사람 당 수송체를 이성질적으로 많은 양 생산하려 할 때 유용하게 사용되는, baculovirus 발현 시스템의 숙주세포로서 이용된다. 그러나 T. ni 세포에 존재하는 내재된 당 수송체의 높은 활동은, 발현된 외재적 당 수송체의 수송활성과 같은 직접적 증거 제시에 장애가 된다. 뿐만 아니라 곤충세포에 내재하는 당 수송체계의 특성에 대해서는 밝혀진 바가 거의 없다. 그래서 본 연구에서는 baculovirus 발현 시스템을 보다 잘 활용하기 위해 T. ni 세포의 2dGlc기질 수송에 D-fructose가 미치는 영향을 살펴 보았으며, T. ni 세포에 발현된 사람 당 수송체의 생물학적 활성을 보다 용이하게 검증하기 위해 발현된 수송체를 [$^3H$] cytochalasin B를 이용하여 photolabelling 하였다. 우선 감염되지 않은 세포와 recombinant AcMPV-GTL 감염시킨 T. ni 세포의 2dGlc uptake를 300 mM D-fructose가 있을 때와 없을 때, 그리고 $20{\mu}M$ cytochalasin B가 있을 때와 없을 때의 상황에서 살펴보았다. 감염되지 않은 세포에서의 육탄당 uptake는 D-fructose에 의해 강력하게 억제 되었으나 cytochalasin B에 의해서는 단지 미미한 억제 효과만을 보여주었다. 흥미롭게도 AcMPV-GTL 바이러스 감염된 T. ni 세포에서는 비록 2dGlc uptake율은 감염되지 않은 세포와 비교해 다소 낮았지만 육탄당 수송 억제 반응은 근본적으로 동일함을 보여 주었다. 또한 [$^3H$] cytochalasin B를 이용한 발현단백질 photolabelling에서는, L-glucose가 존재하는 상황 하에만 하나의 날카롭게 표지된 peak가, 바이러스 감염된 세포에서 관찰되었다. 감염되지 않은 세포에서는 이러한 peak는 관찰되지 않았다. 게다가 D-glucose 존재 하에서는 발현된 단백질의 photolabelling이 완전히 억제되어짐을 보여주어, labelling의 입체선택성(stereoselectivity)을 입증하였다.

• 한국약용작물학회지
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• 제19권2호
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• pp.103-110
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• 2011

This study was performed to enhance anticancer activities of Lithospermum erythrorhizon by eluting high amount of shikonin through ultra high pressure process. Extraction yield was increased up to 5~10% by ultra high pressure process, compare to the normal extraction processes such as water solvent extraction, 70% ethyl alcohol solvent extraction. The cytotoxicity of the extracts ($1.0{\mu}g/m{\ell}$) from ultra high pressure process was showed the lowest cytotoxicity 13.4% for human lung cell (HEL299). The anticancer activities showed 80~85% by adding $1.0{\mu}g/m{\ell}$ of the extracts from ultra high pressure process in several cancer cell lines such as AGS, Hep3B, MCF-7 and HeLa cells. Among them, MCF-7 cell of the endocrine system was highest inhibited than other cells. The anticancer activities of the extracts from ultra high pressure extraction process showed 10~15%, which was higher than the extracts from normal extraction processes. From HPLC analysis of the extracts, the contents of shikonin in the extracts from ultra high pressure process was 11.42% (w/w), which was 20% higher than others. This results indicate that ultra high pressure process could increase the extraction yield of shikonin and other contents, which resulted in higher anticancer activities.

• 동의생리병리학회지
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• 제31권3호
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• pp.159-166
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• 2017

Through this study, the authors investigated the anti-steatosis effects of the Amomum cardamomum ethyl acetate fraction in free fatty acids (FFAs)-induced human hepatocellular carcinoma HepG2 cells. The ethyl acetate fraction of Amomum cardamomum (ACEA) was extracted with 70% ethanol and then the extract was evaporated using a rotary evaporator prior to sequential fractionation. Human hepatocellular carcinoma were treated with different concentrations of ACEA in the presence and absence of FFAs. To demonstrate the reactive oxygen species (ROS) scavenging activity, DCFDA level was analyzed by using in vitro assay system. Cell viability, lipid accumulation, intracellular triglycerides, malondialdehyde (MDA), liver steatosis related signaling molecules and inflammatory cytokines such as interleukin (IL)-6, 8, tumor necrosis factor-alpha ($TNF-{\alpha}$) were also investigated. As results, ACEA inhibited the FFAs-induced ROS, lipid accumulation, intracellular triglycerides, and MDA in a dose dependent manner. Treatment of human hepatocellular cells with ACEA induced the phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK) and carnitine palmitoyltransferase I (CPT1) expression using western blot analysis. ACEA also potently suppressed the FFAs-induced inflammatory cytokines including IL-6, IL-8 and $TNF-{\alpha}$. These results suggest that the ethyl acetate fraction of Amomum cardamoum extract own inhibitory effects of liver steatosis by inhibiting ROS, lipid accumulation, intracellular triglycerides, MDA through AMPK signaling and anti-inflammatory actions.

• BMB Reports
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• 제46권6호
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• pp.322-327
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• 2013

The ATP-binding cassette transporters ABCG5 and ABCG8 form heterodimers that limit absorption of dietary sterols in the intestine and promote cholesterol elimination from the body through hepatobiliary secretion. To identify cis-regulatory elements of the two genes, we have cloned and analyzed twenty-three evolutionary conserved region (ECR) fragments using the CMV-luciferase reporter system in HepG2 cells. Two ECRs were found to be responsive to the Liver-X-Receptor (LXR). Through elaborate deletion studies, regions containing putative LXREs were identified and the binding of $LXR{\alpha}$ was demonstrated by EMSA and ChIP assay. When the LXREs were inserted upstream of the intergenic promoter, synergistic activation by $LXR{\alpha}/RXR{\alpha}$ in combination with GATA4, $HNF4{\alpha}$, and LRH-1, which had been shown to bind to the intergenic region, was observed. In conclusion, we have identified two LXREs in ABCG5/ABCG8 genes for the first time and propose that these LXREs, especially in the ECR20, play major roles in regulating these genes.

• 대한한방내과학회지
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• 제20권1호
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• pp.133-142
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• 1999

To clarifiy the effects of Scolopendrae corpus(S-C) on turmor promotion in two-stage carcinogenesis in mice was investigated. In vivo system, S-C were seen to gave an inhibitory activity on TPA-induced mouse ear edema. In addition, the S-C were proved to have antitumor-promoting activity in two-stage mouse skin carcinogenesis induced by DMBA and two-stage mouse lung carcinogenesis induced by 4-NQO as a initiator plus TPA and glycerol as a promoter. Moreover, S-C significantly exhibited an cytolytic effect in $HepG_2$ cells and showed significant antitumor activity against Sarcoma-180 bearing mice by oral administration. These results suggest that S-C could be effective in adjuvant chemotherapy for human cancer.

• Journal of Pharmaceutical Investigation
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• 제40권6호
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• pp.357-362
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• 2010

To enhance therapeutic effects of insulin-sensitizing adipokine, ADN gene and potent agonists, rosiglitazone for the $PPAR{\gamma}$, cationic liposomes as non-viral vectors were formulated. The particle size and zeta potential of drug loaded and unloaded cationic liposomes were investigated. The complex formation between cationic liposomes and negatively charged plasmid DNA was confirmed and the protection from DNase was observed. In vitro transfection was investigated in HepG2, HeLa, and HEK293 cells by mRNA expression of ADN. Encapsulation efficacy of rosiglitazone-loaded liposomes was determined by UV detection. Particle sizes of cationic liposomes were in the range of 110-170 nm and those of rosiglitazone-loaded cationic liposomes were in the range of 130-180 nm, respectively. Gel retardation of complexes indicated that the complex was formed at weight ratios of cationic lipid to plasmid DNA higher than 20:1. Both complexes protected plasmid DNA from DNase either drug free or drug loading. Encapsulation efficiency of rosiglitazone-loaded emulsion was increased by drug dose. The mRNA expression levels of ADN were dose-dependently increased in cells transfected with plasmid DNA. Therefore, cationic liposomes could be potential co-delivery system for drug and gene.

• Bulletin of the Korean Chemical Society
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• 제34권5호
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• pp.1407-1413
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• 2013

A new compound, perforaphenonoside A (1), along with 11 known compounds (2-12) were isolated from a methanol extract of adventitious roots of Hypericum perforatum. Their chemical structures were elucidated using chemical and physical methods as well as comparison of NMR and mass spectral data with previously reported data. Their inhibition of NF-${\kappa}B$ and activation of PPAR was measured in HepG2 cells using a luciferase reporter system. Among the compounds 3, 6, 7 and 12 inhibited NF-${\kappa}B$ activation stimulated by TNF${\alpha}$ in a dose-dependent manner, with $IC_{50}$ values ranging from 0.85 to $8.10{\mu}M$. Moreover, compounds 1-3, 7, 11 and 12 activated the transcriptional activity of PPARs in a dose-dependent manner, with $EC_{50}$ values ranging from 7.3 to $58.7{\mu}M$. The transactivational effects of compounds 1-3, 7, 11 and 12 were evaluated on three individual PPAR subtypes. Among them, compound 2 activated $PPAR{\alpha}$ transcriptional activity, with 153.97% stimulation at $10{\mu}M$, while compounds 1, 2 and 11 exhibited transcriptional activity of $PPAR{\gamma}$, with stimulation from 124.76% to 126.91% at $10{\mu}M$.

• Journal of Ginseng Research
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• 제44권2호
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• pp.215-221
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• 2020

Background: Panax ginseng has been used for a variety of medical purposes in eastern countries for more than two thousand years. From the extensive experiences accumulated in its long medication use history and the substantial strong evidence in modern research studies, we know that ginseng has various pharmacological activities, such as antitumor, antidiabetic, antioxidant, and cardiovascular system-protective effects. The active chemical constituents of ginseng, ginsenosides, are rich in structural diversity and exhibit a wide range of biological activities. Methods: Ginsenoside constituents from P. ginseng flower buds were isolated and purified by various chromatographic methods, and their structures were identified by spectroscopic analysis and comparison with the reported data. The 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H- tetrazolium bromide method was used to test their cytotoxic effects on three human cancer cell lines. Results: Six ginsenosides, namely 6'-malonyl formyl ginsenoside F₁ (1), 3β-acetoxyl ginsenoside F₁ (2), ginsenoside Rh₂₄ (6), ginsenoside Rh₂₅ (7), 7β-hydroxyl ginsenoside Rd (8) and ginsenoside Rh₂₆ (10) were isolated and elucidated as new compounds, together with four known compounds (3-5 and 9). In addition, the cytotoxicity of these isolated compounds was shown as half inhibitory concentration values, a tentative structure-activity relationship was also discussed based on the results of our bioassay. Conclusion: The study of chemical constituents was useful for the quality control of P. ginseng flower buds. The study on antitumor activities showed that new Compound 1 exhibited moderate cytotoxic activities against HL-60, MGC80-3 and Hep-G2 with half inhibitory concentration values of 16.74, 29.51 and 20.48 μM, respectively.

• Journal of Microbiology
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• 제43권spc1호
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• pp.110-117
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• 2005

Salmonella enterica serovar Typhimurium causes human gastroenteritis and a systemic typhoid-like infection in mice. A critical virulence determinant of Salmonella is the ability to invade mammalian cells. The expression of genes required for invasion is tightly regulated by environmental conditions and a variety of regulatory genes. The hilA regulator encodes an OmpR/ToxR family transcriptional regulator that activates the expression of invasion genes in response to both environmental and genetic regulatory factors. Work from several laboratories has highlighted that regulation of hilA expression is a key point for controlling expression of the invasive phenotype. A number of positive regulators of hilA expression have been identified including csrAB, sirA/barA, pstS, hilC/sirC/sprA, fis, and hilD. HilD, an AraC/XylS type transcriptional regulator, is of particular importance as a mutation in hilD results in a 14-fold decrease in chromosomal hilA::Tn5lacZY-080 expression and a 53-fold decrease in invasion of HEp-2 cells. It is believed that HilD directly regulates hilA expression as it has been shown to bind to hilA promoter sequences. In addition, our research group, and others, have identified genes (hilE, hha, pag, and lon) that negatively affect hilA transcription. HilE appears to be an important Salmonella-specific regulator that plays a critical role in inactivating hilA expression. Recent work in our lab has been directed at understanding how environmental signals that affect hilA expression may be processed through a hilE pathway to modulate expression of hilA and the invasive phenotype. The current understanding of this complex regulatory system is reviewed.

• 한국약용작물학회지
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• 제12권3호
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• pp.243-248
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• 2004

인간 면역 세포 중 하나인 T세포에 대한 각 시료의 생육촉진 활성은 시료의 1.0 g/l에서 기준물질로 사용한 유향열매의 정유 성분이 T세포의 생육촉진 활성 측정에서 가장 좋은 활성을 나타내었으며, 노간주나무의 열매의 정유성분이 줄기 부분의 정유 성분보다 높은 생육 촉진 활성을 보였다. T 세포의 각 cell 당 $TNF-{\alpha}$의 분비량을 살펴보면 모든 시료의 정유 성분들이 배양 2일째 최대분비량을 나타내었고, IL-6의 경우는 모든 시료들이 배양 시간 의존적으로 증가하는 경향을 나타내었다. 유방암 세포인 MCF-7에 대한 각 시료의 생육 억제 활성은 각 정유 성분에 대하여 농도 의존적인 경향을 나타내었으며, 가장 좋은 활성을 나타낸 시료는 노간주나무 열매의 정유 성분으로 약 84%의 높은 생육 억제 활성을 나타내었으며, 유향 열매와 노간주나무 줄기의 정유 성분들도 약 80%에 가까운 높은 생육 억제 활성을 나타내었다. 간암 세포인 Hep3B에 대한 각 정유 성분들의 암세포 생육 억제 활성도 유방암 세포와 같이 각 정유 성분에 대하여 농도 의존적인 경향을 나타내었으며, 유향 열매의 정유 성분이 약 83%의 높은 생육 억제 활성을 나타내어 가장 좋은 활성을 나타내었다. 항암 및 면역기능의 역할에 대한 분자수준에서의 검증을 위해 HL-60 세포의 분화도를 살펴본 결과, 배양 시간 의존적으로 분화도가 증가하는 경향을 나타내었으며, 유향열매의 정유 성분이 Control 군에 비하여 1.38배의 증진활성을 나타내어 가장 높은 분화도를 나타내었으며, 노간주 열매의 정유 성분이 줄기의 정유 성분에 비하여 높은 분화도를 나타내었다.