• Molecules and Cells
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• 제38권4호
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• pp.327-335
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• 2015

Piperlongumine, a natural alkaloid isolated from the long pepper, selectively increases reactive oxygen species production and apoptotic cell death in cancer cells but not in normal cells. However, the molecular mechanism underlying piperlongumine-induced selective killing of cancer cells remains unclear. In the present study, we observed that human breast cancer MCF-7 cells are sensitive to piperlongumine-induced apoptosis relative to human MCF-10A breast epithelial cells. Interestingly, this opposing effect of piperlongumine appears to be mediated by heme oxygenase-1 (HO-1). Piperlongumine upregulated HO-1 expression through the activation of nuclear factor-erythroid-2-related factor-2 (Nrf2) signaling in both MCF-7 and MCF-10A cells. However, knockdown of HO-1 expression and pharmacological inhibition of its activity abolished the ability of piperlongumine to induce apoptosis in MCF-7 cells, whereas those promoted apoptosis in MCF-10A cells, indicating that HO-1 has anti-tumor functions in cancer cells but cytoprotective functions in normal cells. Moreover, it was found that piperlongumine-induced Nrf2 activation, HO-1 expression and cancer cell apoptosis are not dependent on the generation of reactive oxygen species. Instead, piperlongumine, which bears electrophilic ${\alpha},{\beta}$-unsaturated carbonyl groups, appears to inactivate Kelch-like ECH-associated protein-1 (Keap1) through thiol modification, thereby activating the Nrf2/HO-1 pathway and subsequently upregulating HO-1 expression, which accounts for piperlongumine-induced apoptosis in cancer cells. Taken together, these findings suggest that direct interaction of piperlongumine with Keap1 leads to the upregulation of Nrf2-mediated HO-1 expression, and HO-1 determines the differential response of breast normal cells and cancer cells to piperlongumine.

• Journal of Periodontal and Implant Science
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• 제34권3호
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• pp.607-622
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• 2004

Recently epidemiologic studies have indicated that the patients with periodontitis may have increased risk of ischemic cardiovascular events, and have suggested the important roles of blood cytokines and acute reactant proteins in the systemic infection and inflammatory response. Periodontitis and coronary heart disease (CHD) may share the common risk factors and the genetic mechanism associated with interleukin(IL)-1A, B and RA genotype may be involved in the production of IL-1. This study was aimed to investigate the relationship between angiographically defined CHD and periodontitis as chronic Gram-negative bacterial infection and to determine whether the IL-1 gene polymorphism is associated in both diseases. Patients under the age of 60 who had undergone diagnostic coronary angiography were enrolled in this study. Subjects were classified as positive CHD (+CHD, n=37) with coronary artery stenosis more than 50% in at least one of major epicardial arteries, and negative CHD (-CHD, n=30) without significant stenosis. After recording the number of missing teeth, periodontal disease severity was measured by means of plaque index (PI), gingival index (GI), bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and radiographic bone loss around all remaining teeth. Gingival crevicular fluid (GCF) was collected from the 4 deepest periodontal pockets and assessed for cytokine ($IL-1{\beta}$, IL-6, IL-1ra, tumor necrosis $factor-{\alpha}$, and prostaglandin $E_2$). Additionally, blood CHD markers, lipid profile, and blood cytokines were analyzed. IL-1 gene cluster genotyping was performed by polymerase chain reaction and enzyme restriction using genomic DNA from buccal swab, and allele 2 frequencies of IL-1A(+4845), IL-1B(+3954), IL-B(-511), and IL-1RA(intron 2) were compared between groups. Even though there was no significant difference in the periodontal parameters between 2 groups, GCF level of $PGE_2$ was significantly higher in the +CHD group(p<0.05). Correlation analysis showed the positive relationship among PD, CAL and coronary artery stenosis(%) and blood $PGE_2$. There was also significant positive relationship between the periodontal parameters (PI, PD, CAL) and the blood CHD markers (leukocyte count, C-reactive protein, and lactic dehyrogenase). IL-1 gene genotyping showed that IL-1A(+3954) allele 2 frequency was significantly higher in the +CHD group compared with the -CHD group (15% vs. 3.3%, OR 5.118,p=0.043). These results suggested that periodontal inflammation is related to systemic blood cytokine and CHD markers, and contributes to cardiovascular disease via systemic inflammatory reaction. IL-1 gene polymorphism might have an influence on periodontal and coronary heart diseases in Korean patients.

• Asian Pacific Journal of Cancer Prevention
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• 제13권4호
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• pp.1457-1461
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• 2012

Purpose: Non small cell lung cancer (NSCLC) is the leading worldwide source of cancer-related deaths. Although some drugs targeting EGFR mutations have been developed, most advanced cases are still incurable. New targets for anticancer drugs are demanded. The kringle 1 domain of hepatocellular growth factor alpha chain (HGFK1) is a potent anti-angiogenesis factor. It has also emerged as a potential anticancer factor in hepatocellular carcinoma (HCC). The expression of HGFK1 protein in patients with NSCLC has not been reported to date. Method: Here, we assessed HGFK1 expression by Western blotting in 103 cases with advanced NSCLC to investigate the impact of HGFK1 on survival. Results: Results revealed 33 (30.1%) patients were classified as high expressors, this being significantly associated with less remote metastasis (P = 0.002) but not with lymph node metastasis (P = 0.062). There was also a significant association between HGFK1 expression and tumor size (P = 0.025) as well as clinical stage (P = 0.012). Kaplan-Meier survival analysis showed that both overall survival (OS) and progression free survival (PFS) of patients with HGFK1 expression were longer than those of patients without HGFK1 expression (P = 0.004 and P = 0.001 respectively). HGFK1 reversed gefitinib inhibition in the resistent NSCLC cell line A431/GR but did not inhibit the proliferation of NSCLC cells A431 and A431/GR directly. Reversion of gefitinib inhibition in A431/GR cells by HGFK1 was related to decreased phosphorylation of ERK and STAT5. Conclusions: HGFK1 may be a useful prognostic factor of advanced NSCLC patients and a potential drug for gefitinib resistant patients.

• 한국양식학회지
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• 제9권4호
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• pp.353-361
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• 1996

자주복 유어의 필수지방산 요구 및 사료중의 적정함량을 규명하기 위하여, 평균체중 3.45g의 유어를 사용하여 사육실험을 실시하였다. 8주간의 사육실험의 결과, 성장은 실험개시 4주부터 실험구들 사이에 차이가 보여 실험 종료시에는 $0.5\~1\%$ n-3 HUFA 실험구와 $0.5\%$ DHA 실험구가 다른 실험구에 비교하여 우세한 성장을 보였다(P<0.05). 또한, EPA, DHA 및 n-3 HUFA 실험구 사이에 효율성의 비교에서 EPA$1\%$ EPA실험구는 사료중에 $0.5\%$에서 $1\%$로 지방산을 증량시킴에 의해 첨가효과가 인정되었으나, DHA와 n-3 HUFA실험구는 EPA실험구와 상이하게 $1\%$ DHA실험구에서 급격한 성장의 저하와 $1\%$ n-3 HUFA 실험구에서는 성장의 정체가 나타났다. 실험기간동안에 생잔률은 $73\~93\%$의 범위로 지방산과의 관련성은 보이지 않았다. 사료효율은 n-3 HUFA실험구와 $0.5\%$ DHA실험구에서 높았고, $1\%$ LNA실험구는 $0.5\~1\%$ EPA실험구와 $0.3\%$ DHA실험구와의 차이가 거의 없었다. 이상의 결과로 부터, 유어기의 자주복은 EPA나 DHA등의 n-3 HUFA를 필수지방산으로 요구함이 사료되며, 사료중의 $0.5\%$ 정도의 HUFA 또는 $0.5\%$ DHA가 적정함량으로 추측된다.

• 한국식품과학회지
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• 제41권3호
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• pp.313-319
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• 2009

본 실험은 해수에서 배양된 Spirulina maxima를 $100^{\circ}C$와 초음파 병행 $60^{\circ}C$ 물 추출물, $80^{\circ}C$ EtOH 추출물을 이용하여 실험을 수행하였다. AOAC법에 의한 일반 조성 및 무기질, 아미노산 조성 분석결과 조 단백질과 무기질 중 나트륨이 각각 56, 60%의 비교적 높은 비율을 차지했으며, 다양한 균주의 필수 아미노산을 포함하고 있었다. 그중 피부와 관련된 leucine의 함량이 9.83%로 상대적으로 풍부하여 S. maxima 피부면역 활성 증진에 효율적인 소재의 가능성을 확인했다. 본 연구에서는 피부 면역과 인체 면역과의 밀접한 상관성이 있으므로 본 연구에서는 T 세포와 B 세포의 생육증진 및 cytokine의 분비량 측정과 NK 세포의 활성 및 항염증 효과인 hyaluronidase 저해 활성 효과를 측정하였다. 먼저 정상세포(HEK293)에 대한 세포독성 결과 인체에 큰 독성을 나타내지 않는 것으로 나타난 S. maxima를 추출조건에 따른 면역 증진 효과를 비교한 결과 $60^{\circ}C$ 초음파 병행 물 추출물이 면역 B세포와 T세포에서 각각 $11.3{\times}10^4$ cells/mL, $12.8{\times}10^4$ cells/mL으로 6일째 가장 높은 생육도를 보였다, 이러한 면역 세포들이 분비하는 cytokine(IL-6, TNF-${\alpha}$)의 분비량은 가장 높은 생육도를 보인 $60^{\circ}C$ 초음파 병행 물 추출물을 첨가한 B세포, T세포의 경우 IL-6와 TNF-${\alpha}$는 유의적으로 증가된 분비량을 보이며, 면역 및 기능성 소재로서의 활성이 뛰어난 것으로 나타났다. B세포에 각 시료를 첨가하여 배양 후 그 배양액을 NK 세포에 첨가하였을 때, 6일 동안 생육도를 관찰하였는데 시료에 대한 생육도가 증가하며, 6일째 $12.40{\times}10^4$ cells/mL를 나타내어 $11.49{\times}10^4$ cells/mL를 나타낸 대조군에 비해 높은 활성을 나타내었으며, 이로써 활성물질을 포함하고 있음을 알 수 있다. 또한 항염증 효과의 hyaluronidase 저해 활성 효과에서 $60^{\circ}C$ 초음파 병행 물 추출물이 1.0 mg/mL 농도일 때 62.4%의 저해 활성을 보여 $100^{\circ}C$ 이상의 고온 추출에서 야기되는 단백질이나 미립자 활성 물질의 파괴가 최소화되고 저온 초음파 추출 시 유용성분들의 용출량 증가와 새로운 물질들의 생산 혹은 활성변형에 의한 추출을 통해 인체면역과 피부면역 활성 증진에 활용이 가능한 효율적인 소재화가 가능한 것을 확인했다. 이로써, 본 연구 논문은 S. maxima의 물질 분리보다 피부면역관련 활성을 통한 소재로서의 탐색이 주목적이므로 향후 이 추출물의 분리, 동정을 통해 피부 면역 활성도에 따라 물질의 구조 분석에 관한 연구를 보이고자 한다.

• 대한본초학회지
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• 제27권6호
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• pp.115-122
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• 2012

Objectives : This study evaluated activities and ingredient contents concerning extracts according to extraction solvents of Insampaedok-san (IS, Renshen bai du-san). Methods : The herbal constituents of IS were extracted with water and 70% ethanol at $100^{\circ}C$ for 2 hr. Using the HPLC system, the six ingredient contents of different solvent extracts of IS were analyzed. The nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$) production and proinflammatory cytokines were measured in RAW264.7 cells stimulated with lipopolysaccharide (LPS). The macrophage-derived chemokine (MDC/CCL22) and regulated on activation normal T-cell expression and secreted (RANTES/CCL5) production were measured in HaCaT and BEAS-2B cells stimulated tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interferon-${\gamma}$ (IFN-${\gamma}$). The activities of glycerol-3-phosphate dehydrogenase (GPDH) and leptin level were measured in 3T3-L1 cells. Results : The calibration curves showed good linearity ($r^2$=1.0000) for different concentration ranges. The contents of liquiritin, naringin, hesperidin, neohesperirin and glycyrrizin in 70% ethanol extracts of IS were relatively higher than that of water extract, however the content of ferulic acid in 70% ethanol and water extract of IS were similar. The extraction solvents of water and 70% ethanol were evaluated inhibitory effect on the production of NO, $PGE_2$, TNF-${\alpha}$ and IL-6 in RAW 264.7 cells. Their extractions were inhibitory effect on production of MDC/CCL22 and RANTES/CCL5 in HaCaT cell and BEAS-2B cell, respectively. In addition, evaluated reduced on GPDH activity and leptin level in 3T3-L1 preadipocyte cell. Conclusions : Our results suggest that IS extracts were inhibitory effects of disease such as inflammation, allergies and obesity.

• The Korean Journal of Physiology and Pharmacology
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• 제15권1호
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• pp.61-66
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• 2011

P2Y receptors are metabotropic G-protein-coupled receptors, which are involved in many important biologic functions in the central nervous system including retina. Subtypes of P2Y receptors in retinal tissue vary according to the species and the cell types. We examined the molecular and pharmacologic profiles of P2Y purinoceptors in retinoblastoma cell, which has not been identified yet. To achieve this goal, we used $Ca^{2+}$ imaging technique and western blot analysis in WERI-Rb-1 cell, a human retinoblastoma cell line. ATP ($10\;{\mu}M$) elicited strong but transient $[Ca^{2+}]_i$ increase in a concentration dependent manner from more than 80% of the WERI-Rb-1 cells (n=46). Orders of potency of P2Y agonists in evoking $[Ca^{2+}]_i$ transients were 2MeS-ATP>ATP>>UTP=${\alpha}{\beta}$-MeATP, which was compatible with the subclass of $P2Y_1$ receptor. The $[Ca^{2+}]_i$ transients evoked by applications of 2MeS-ATP and/or ATP were also profoundly suppressed in the presence of $P2Y_1$ selective blocker (MRS 2179; $30\;{\mu}M$). $P2Y_1$ receptor expression in WERI-Rb-1 cells was also identified by using western blot. Taken together, $P2Y_1$ receptor is mainly expressed in a retinoblastoma cell, which elicits $Ca^{2+}$ release from internal $Ca^{2+}$ storage sites via the phospholipase C-mediated pathway. $P2Y_1$ receptor activation in retinoblastoma cell could be a useful model to investigate the role of purinergic $[Ca^{2+}]_i$ signaling in neural tissue as well as to find a novel therapeutic target to this lethal cancer.

• 한국방사선학회논문지
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• 제5권6호
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• pp.401-407
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• 2011

TE를 변화시킨 정상인 대뇌의 MR spectrum에서 주요 대사물질의 면적과 SNR을 측정하여 PRESS 펄스파형과 STEAM 펄스파형 그리고 1.5T와 3.0T간의 자장세기에 따른 spectrum 간의 차이를 알아보고자 하였다. Phantom 실험을 통하여 적절한 TR을 정한 후, 정상인 지원자 10명(3.0T 5명, 1.5T 5명 ; 남 22~30세 : 평균 26세)을 대상으로 단일용적기법의 STEAM과 PRESS 기법을 시행하였다. 사용된 장비는 3.0T MR scanner(Magnetom Trio, SIEMENS, Germany)와 1.5T MR scanner(Signa Twinspeed GE, USA)이였다. 영상변수는 TR은 2000ms, TE는 30ms, 40ms, 50ms, 60ms, 90ms, 144ms, 288ms, NA는 96, 용적 크기는 $20{\times}20{\times}20mm3$로 하였으며, spectrum 획득시간은 3분 20초였다. 획득한 데이터는 후처리과정을 통하여 PRESS와 STEAM, 그리고 1.5T와 3.0T system 간의 NAA, Cho, Cr 등의 단순면적값과 SNR을 비교하였다. 또한 육안적 관찰을 통하여 각 대사물질들의 관찰정도를 비교하였다. 1.5T와 3.0T MR spectrum을 분석한 결과, STEAM과 PRESS의 주요 대사물질의 단순 면적값과 SNR은 TE가 증가함에 따라 감소하는 경향을 보였으며, PRESS는 STEAM보다 1.5T에서 1.4배, 3.0T에서 1.3배 높은 SNR을 보였다. 자장의 세기에 따른 SNR 비교에서는 TE가 30ms에서 3.0T가 1.5T보다 약 2배 정도 높은 SNR을 보였으나 TE값이 증가함에 따라 3.0T에서의 SNR 감소율이 1.5T에서의 SNR 감소율보다 커서 TE가 90ms 이상부터는 큰 차이가 없었다. 반면 3.0T의 spectrum에서는 1.5T에서 구분할 수 없었던 ${\alpha}$-Glx, ${\beta}{\cdot}{\gamma}$-Glx, NAA complex등 작은 대사물질들을 보다 정확히 감별 할 수 있었고 short TE의 PRESS일 때 short TE의 STEAM보다 작은 대사물질들이 잘 관찰 되었다. 3.0T spectrum의 해상도와 SNR이 1.5T spectrum에 비하여 우수함을 알 수 있었다. 그러나 90ms이상의 long TE에서는 3.0T와 1.5T spectrum간의 SNR은 차이가 없었다. 따라서 고자장하에서의 자기공명분광법은 30ms 이하의 짧은 TE를 이용한 PRESS 펄스 파형을 사용하는 것이 임상적으로 유용하게 사용될 수 있을 것이라 사료된다.

• 한국작물학회지
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• 제52권4호
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• pp.447-452
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• 2007

괴경 내부에 적색과 보라색 안토시아닌 색소가 풍부하게 함유되어 있고, 색상의 기호도 및 건강 기능성으로 인해 소비자로부터 기호도가 증대된 유색감자의 생리적 활성을 검토하기 위해 유색감자의 대조구로 괴경 내부가 백색인 수미 품종을 사용하고, 유색감자로서 괴경 내부가 적색인 대관 1-102호, 괴경 내부가 보라색인 자심품종과 대관 1-104호로 구분하여 항산화, 활성산소 소거능 및 항고혈압 활성을 비교, 검토한 결과는 아래와 같다. 1. 유색감자 추출물의 항산화 활성 검정결과 추출물의 농도가 $10{\mu}g/mL$ 수준에서는 유색감자별 차이를 나타내지 않았으나, $50{\mu}g/mL$을 처리한 경우 시료 간 차이를 뚜렷하게 나타내었으며, 유색감자 중 괴경내부의 색상이 적색인 대관 1-102호가 34%의 높은 저해율을 나타내어 30%의 저해율을 나타낸 활성평가 대조구인 $\alpha-tocopherol$보다 높은 항산화 활성을 나타내었다. 2. 유색감자 추출물의 자유 라디칼 소거 활성에서는 괴경 내부가 보라색인 대관 1-104호가 50%의 소거능을 나타내는 농도가 $9.83{\mu}g/mL$로 가장 높은 라디칼 소거활성을 나타내었으며, 이 결과 또한 대조시약인 $\alpha-tocopherol$이나 BHT보다 높은 소거 활성을 나타내었다. 3. 유색감자 추출물의 xanthine oxidase 활성 억제효과를 검토한 결과 유색감자 간에 큰 차이를 나타내어 대관 1-102호와 1-104호의 경우 xanthine oxidase 활성을 50% 저해하는 데 필요한 농도는 약 $26{\mu}g/mL$ 수준으로 가장 높게 조사되었으며, 다음으로 수미($46{\mu}g/mL$), 자심($49{\mu}g/mL$)의 순으로 조사되었다. 4. 유색감자 추출물의 ACE 저해 활성을 검토한 결과 항고혈압 대조구 활성물질인 captopril의 수준에는 미치지 못하나 유색감자 간에는 명확한 차이를 나타내었다. 괴경내부색상이 적색인 대관 1-102호가 $117.7{\mu}g/mL$으로 가장 높은 ACE 저해 활성을 나타내었으며, 내부색상이 보라색인 대관 1-104호와 자심은 $130{\mu}g/mL$로 유사한 양상을 나타내었으며, 괴경내부 색상이 백색을 나타내는 수미품종은 $192{\mu}g/mL$으로 가장 낮은 활성을 나타내었다. 5. 이상의 결과를 종합해 볼 때 괴경 내부에 적색 혹은 보라색 안토시아닌을 다량 함유한 유색감자는 기존 백색을 가진 일반감자에 비해 항산화, 자유레디칼 소거활성 및 항고혈압 활성이 높고, 시각적 식미감을 증대시키므로 유색감자는 기능성이 증대된 식용감자로서의 이용가치가 충분한 것으로 판단된다.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.193-213
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• 1995

The purpose of this study is to investigate a possible contribution of nonspecific esterases, which occur in the oral cavity, to the degradation of ester bonds in polymethacrylates. One of the problems connected with the use of composite resins for restorations is their inadequate resistance to wear. It has been shown that methacrylate hydrolysis can be catalyzed by enzymes and that a carboxylic hydrolase (porcine liver esterase) catalyzed the hydrolysis of several mono - and dimethacrylates. The softening effect on a BISGMA/TEGDMA polymer induced by hydrolase will accelerate the in vivo wear of the polymer. Porcine liver esterase (EC 3.1.1.1) 3.2 mol/L $(NH_4)_2$ $SO_4$ was obtained from Sigma Chemical Company. The esterase activity of one unit is defined as the amount of enzyme capable of hydrolyzing $l{\mu}mol$ ethyl butyrate per min at pH 8.0 AT $25^{\circ}C$. Phosphate buffer, 10mmol/L, pH 7.0, was made by adjustment of a solution of $Na_2HPO_4$ with $H_3PO_4$. Composite resins used in this study are Silux Plus, Z-100, Durafil VS, and Prisma APH. Cylindrical specimens, 14mm in diameter and 3mm thick, of Silux Plus, Z-100, Durafil VS, Prisma APH were polymerized under the celluloid strip. 60 specimens were divided into 2 groups. One group was emersed only in buffer solution, the other group was emersed in buffer and enzyme solution. Silux Plus and Z-100 were divided into 2 subgroups, one subgroup was cured only Visilux 2. And the other subgroup was cured Visilux 2 and Triaid II. Thereafter, specimens were polished to its best achievable surface according to manufacture's directions. The Vickers hardness of the specimens was measured after 1, 2, 4, 7, 9, 15, 50 days. The solutions were changed after each measurement. Composite resin surfaces were evaluated for the surface roughness with profilometer (${\alpha}$-step 200, Tencor instruments, USA) after 1 and 50 days. And then surfaces of specimens were pictured with stereosopy after 1 and 50 days. The results were as follows. 1. The surface hardness of Silux plus, durafil VS, and Prisma APH were decreased with time. But, the surface hardness of Z-100 was not decreased. 2. The surface hardness of all composite resins was decreased by esterase. 3. Composite resins, which were light-cured by Visilux 2 and concomitantly baked by oven, showed more hardened surface than light-cured by Visilux 2 only. 4. Significant surface changes were occured in Silux plus after esterase treatment.