• Archives of Pharmacal Research
• /
• 제29권10호
• /
• pp.911-920
• /
• 2006

Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

• 대한본초학회지
• /
• 제20권3호
• /
• pp.29-41
• /
• 2005

Objectives : The use of natural products with therapeutic properties is as ancient as human civilisation and, for a long time, mineral, plant and animal products were the main sources of drugs. Catalposide, the major iridoid glycoside isolated from the stem bark of Catalpa ovata G. Don (Bignoniceae) has been shown to possess anti-microbial and anti-tumoral properties. Heme oxygenase-1 (HO-1) is a stress response protein and is known to play a protective role against the oxidative injury. In this study, we examined whether catalposide could protect Neuro 2A cells, a kind of neuronal cell lines, from oxidative damage through the induction of HO-1 protein expression and HO activity. We also examined the effects of catalposide on the productions of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and nitric oxide (NO) on RAW 264.7 macrophages activated with the endotoxin lipopolysaccharide. Methods : HO-1 expression in Neuro 2A cells was measured by Western blotting analysis. NO and $TNF--{\alpha}$ produced by RAW 264.7 macrophage were measured by Griess reagent and enzyme-linked immunosorbent assay, respectively. Results : The treatment of the cells with catalposide resulted in dose- and time-dependent up-regulations of both HO-1 protein expression and HO activity. Catalposide protected the cells from hydrogen peroxide-induced cell death. The protective effect of catalposide on hydrogen peroxide-induced cell death was abrogated by zinc protoporphyrin IX, a HO inhibitor. Additional experiments revealed the involvement of CO in the cytoprotective effect of catalposide-induced HO-1. In addition, catalposide inhibited the productions of $TNF--{\alpha}$ and NO with significant decreases in mRNA levels of $TNF--{\alpha}$ and inducible NO synthase. Conclusions : Our results indicate that catalposide is a potent inducer of HO-1 and HO-1 induction is responsible for the catalposide-mediated cytoprotection against oxidative damage and that catalposide may have therapeutic potential in the control of inflammatory disorders.

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

• 한국식품영양과학회지
• /
• 제46권4호
• /
• pp.409-416
• /
• 2017

천연물 유래 물질의 항염증 활성에 대한 잠재성을 평가하기 위한 일환으로 오배자에서 분리한 PGG가 LPS로 자극한 마우스 대식세포인 RAW264.7 세포에서 염증반응에 미치는 영향에 대해 평가하고 관련 메커니즘에 대해 검토하였다. PGG는 LPS 자극에 의해 유도된 iNOS 및 COX-2 단백질 발현량을 감소함으로써 NO와 $PGE_2$ 생성을 억제할 뿐만 아니라 IL-6, TNF-${\alpha}$와 같은 pro-inflammatory cytokine의 분비를 억제하였다. 이러한 효과는 전사인자인 NF-${\kappa}B$의 세포질에서 핵으로의 이동을 억제함으로써 나타나는 것으로 판단된다. 이러한 결과로부터 PGG가 염증 반응을 저해하는 효과가 있는 것으로 나타나 향후 염증성 질환을 예방, 개선 및 치료하는 데 유용한 물질로 사용될 가능성이 있을 것으로 생각된다.

• 대한소아치과학회지
• /
• 제29권3호
• /
• pp.463-468
• /
• 2002

단방성 법랑모세포종(Unicystic ameloblastoma)은 임상적, 방사선적 및 병리학적 소견과 치료방법 등이 법랑모세포종과 구별되어 따로 분류된 질환이다. 임상적, 방사선적으로는 치성낭종의 소견을 보이는 단방정 병소이며 조직학적으로 낭종의 소견과 함께 법랑모세포종의 소견을 보인다. 하악 구치부에 호발하며 악골에 무통성 종창을 야기 할 수 있으나 대개는 무증상인 경우가 많고 대부분 미맹출치 치관을 둘러싼 방사선 투과성 병소로 나타나며 방사선적으로 함치성낭(dentigerous cyst)이나 잔류낭(residual cyst)과 유사하다. 조직학적으로 낭종상피의 법랑모세포종성 변화(luminal type), 낭종강 내 종양결절의 돌출(intraluminal type), 결합조직 내로 법랑모세포종 세포의 증식(mural type) 등의 소견을 보인다. 본 증례는 좌측 안면부 종창을 주소로 내원한 8세 남자 환아에서 임상적, 방사선적 검사 후 낭종의 완전 적출술(enucleation)및 장골이식(iliac bone graft)을 시행하였으며 생검을 통해 단방성 법랑모세포종이라 확진되었고 양호한 치료 결과를 얻었기에 이에 보고하는 바이다.

• IMMUNE NETWORK
• /
• 제13권2호
• /
• pp.70-74
• /
• 2013

L-ascorbic acid (vitamin C) is one of the well-known antiviral agents, especially to influenza virus. Since the in vivo antiviral effect is still controversial, we investigated whether vitamin C could regulate influenza virus infection in vivo by using Gulo (-/-) mice, which cannot synthesize vitamin C like humans. First, we found that vitamin C-insufficient Gulo (-/-) mice expired within 1 week after intranasal inoculation of influenza virus (H3N2/Hongkong). Viral titers in the lung of vitamin C-insufficient Gulo (-/-) mice were definitely increased but production of anti-viral cytokine, interferon (IFN)-${\alpha}/{\beta}$, was decreased. On the contrary, the infiltration of inflammatory cells into the lung and production of pro-inflammatory cytokines, tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-${\alpha}/{\beta}$, were increased in the lung. Taken together, vitamin C shows in vivo antiviral immune responses at the early time of infection, especially against influenza virus, through increased production of IFN-${\alpha}/{\beta}$.

• 대한본초학회지
• /
• 제23권3호
• /
• pp.127-134
• /
• 2008

Objectives : The present study was designed to investigate whether Ostericum koreanum (OK) could regulate lipopolysaccharide (LPS)-induced inflammatory response in vitro and in vivo. Methods : To evaluate of anti-inflammatory effect of OK, we examined Nitric oxide (NO), proinflammatory cytokines production in LPS-stimulated RAW264.7 cells. Furthermore, we checked molecular mechanism especially in the phosphorylation of mitogen-activated protein kinases (MAPKs) and the degradation of inhibitory kappa B a ($Ik-B{\alpha}$) using western blot and also investigated survival of mice in LPS-mediated endotoxin shock. Results : 1. Extract from OK itself have weak cytotoxic effect on RAW264.7 cells. Extract from OK inhibited LPS-induced NO, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, IL-6 and IL-10 production in RAW264.7 cells. 2. OK inhibited the phosphorylation of MAPKs, such as p38, extracelluar signal-regulated kinase (ERK1/2) and c-Jun NH2-terminal kinase (JNK) and also the degradation of $I{\kappa}-B{\alpha}$ in the LPS-stimulated RAW264.7 cells 3. OK did not inhibit LPS-induced endotoxin shock. Conclusions : OK down-regulated LPS-induced NO and cytokines production through suppressing activation of MAPKs and degradation of $I{\kappa}-B{\alpha}$. Our results suggested that OK may be a beneficial drug against inflammatory diseases.

• 한국식품과학회지
• /
• 제44권1호
• /
• pp.82-88
• /
• 2012

감국 에탄올 추출물이 $H_2O_2$로 유도한 산화적 스트레스 상황에서 MC3T3-E1 조골세포의 증식 및 분화, ROS 생성 및 염증 매개성 cytokine인 TNF-${\alpha}$ 생성 등에 미치는 영향을 분석하였다. $H_2O_2$로 유도한 산화적 스트레스 상황에서 감국 에탄올 추출물은 30-100 ${\mu}g/mL$ 농도 범위에서 조골세포의 증식을 유의적으로 증가시켰다. 또한, 감국 에탄올 추출물 200 ${\mu}g/mL$ 농도에서 ALP 활성이 약 1.5배 유의적인 증가를 나타냈다. 그러나 collagen 합성에는 유의적 차이를 보이지 않았다. Mineralization 측정에서는 200 ${\mu}g/mL$ 농도에서 대조군에 비해 유의적 증가를 보였다. $H_2O_2$로 유도한 산화적 스트레스 상황에서 감국 에탄올 추출물이 intracellular ROS 생성에 미치는 영향을 측정해본 결과, 30 ${\mu}g/mL$ 농도에서 antioxidant인 trolox 20 ${\mu}M$과 유사한 ROS 생성수준을 나타내어 유사한 항산화 효과를 보였으며, 그 이상의 농도에서는 더 높은 항산화 효과를 나타냈다. $H_2O_2$로 유도한 산화적 스트레스 상황에서 조골세포의 collagen 및 ALP의 합성을 억제하고 파골세포로의 분화 증강과 골흡수를 촉진시키는 것으로 알려진 TNF-${\alpha}$ 생성정도를 측정한 결과, 감국 에탄올 추출물 처리에 의해 농도 의존적으로 생성이 감소되었으며, 200 ${\mu}g/mL$ 농도에서 대조군 대비 89% 수준을 나타내어 유의적 차이를 보였다. 이상의 결과를 통해 감국 에탄올 추출물은 $H_2O_2$로 유도된 산화적 스트레스 상황에서 세포 내 ROS 생성과 염증매개 cytokine인 TNF-${\alpha}$ 생성을 감소시킴으로써 조골세포 손상과 활성 감소를 억제하고, 증식과 분화를 촉진시키는 효과가 있는 것을 확인할 수 있었다. 따라서 감국 에탄올 추출물의 골다공증 예방을 위한 식물성 에스트로젠(phytoestrogen) 및 항산화 소재로의 이용 가능성이 있을 것으로 사료된다.

• Tuberculosis and Respiratory Diseases
• /
• 제70권1호
• /
• pp.21-27
• /
• 2011

Background: Although the gold standard method for research trials on epidermal growth factor receptor (EGFR) mutations has been direct sequencing, this approach has the limitations of low sensitivity and of being time-consuming. Peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping is known to be a more sensitive detection tool. The aim of this study was to compare the detection rate of $EGFR$ mutation and EGFR-tyrosine kinase inhibitor (TKI) responsiveness according to $EGFR$ mutation status using both methodologies. Methods: Clinical specimens from 112 NSCLC patients were analyzed for $EGFR$ mutations in exons 18, 19, 20, and 21. All clinical data and tumor specimens were obtained from 3 university hospitals in Korea. After genomic DNA was extracted from paraffin-embedded tissue specimens, both PNA-mediated PCR clamping and direct-sequencing were performed. The results and clinical response to $EGFR$-TKIs were compared. Results: Sequencing revealed a total of 35 (22.9%) mutations: 8 missense mutations in exon 21 and 26 deletion mutations in exon 19. PNA-mediated PCR clamping showed the presence of genomic alterations in 45 (28.3%) samples, including the 32 identified by sequencing plus 13 additional samples (6 in exon 19 and 7 in exon 21). Conclusion: PNA-mediated PCR clamping is simple and rapid, as well as a more sensitive method for screening of genomic alterations in $EGFR$ gene compared to direct sequencing. This data suggests that PNA-mediated PCR clamping should be implemented as a useful screening tool for detection of $EGFR$ mutations in clinical setting.

• 동의생리병리학회지
• /
• 제21권4호
• /
• pp.973-981
• /
• 2007

In the present study, we investigated the antiproliferative activity of the water extract of Samgibopae-tang (SGBPT) in NCI-H460 and A549 non-small-cell lung cancer cell lines. We found that exposure of A549 cells to SGBPT resulted in the growth inhibition in a dose-dependent manner as measured by MTT assay, however SGBPT did not affect the growth of NCI-H460 cells. The antiproliferative effect by SGBPT treatment in A549 cells was associated with morphological changes such as membrane shrinking and cell rounding up. SGBPT treatment did not induce the cell cycle arrest in both cell lines, however the frequency of sub-G1 population was concentration-dependently increased by SGBPT treatment in A549 cells. SGBPT treatment partially induced the expression of tumor suppressor p53 in A549 cells and the expression of cyclin-dependent kinase inhibitor p21(WAF1/CIP1) was markedly increased in both transcriptional and translational levels in A549 cells. The up-regulation of p21 by SGBPT occurred in a similar a concentration dependent manner to that observed with the inhibition of cell viability and induction of sub-G1 population of the cell cycle. However SGBPT treatment did not affect other growth regulation-related genes such as early growth response-1 (Egr-1), nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1), inducible nitric oxide synthease (iNOS), cyclooxygenases (COXs), telomere-regulatory factors in A549 as well as NCI-H460 cells. Taken together, these findings suggested that SGBPT-induced inhibition of human lung carcinoma A549 cell growth was aoosciated with the induction of p21 and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of SGBPT.