• 한국식품영양과학회지
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• 제45권3호
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• pp.445-451
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• 2016

본 연구는 고사리 및 취나물과 같은 산채가 첨가된 된장 및 일반 된장의 면역 활성을 비교하기 위하여 선천면역계의 대표적인 세포인 대식세포와 적응면역계에서 중추적인 역할을 수행하는 비장세포에 각각 산채첨가 된장 추출물(WPDJ) 및 일반 된장(DJ) 추출물을 처리하여 각각의 면역세포의 세포증식률과 사이토카인 분비능에 미치는 영향에 관하여 측정하여 보았다. 대식세포 및 비장세포에 WPDJ 및 DJ를 125, 250 및 $500{\mu}g/mL$의 농도로 처리하였을 때 두 추출물 모두 대식세포 및 비장세포에 대한 세포독성을 유발하지 않았으며, 농도 의존적으로 세포증식률을 증가시키는 것으로 관찰되었다. 대식세포의 사이토카인의 분비능에 관하여 알아본 결과 WPDJ 처리구에서 더 높게 증가하는 것으로 관찰되었다. 또한 마우스 비장에서 유리된 비장세포에 WPDJ 및 DJ를 125, 250 및 $500{\mu}g/mL$의 농도로 처리하였을 때 Th1 type의 사이토카인인 IL-2와 IFN-${\gamma}$의 분비능은 유의적으로 증가한 반면, 알레르기를 유도하는 것으로 알려진 Th2 type의 사이토카인인 IL-4의 생성에는 영향을 미치지 않는 것으로 관찰되었다. 따라서 산채를 첨가하여 제조한 된장은 일반 된장보다 면역 활성을 더 높게 증가시키는 것으로 생각한다.

• The Korean Journal of Physiology and Pharmacology
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• 제21권5호
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• pp.519-529
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• 2017

Sodium butyrate (SB) has various metabolic actions. However, its effect on dipeptidyl peptidase 4 (DPP-4) needs to be studied further. We aimed to evaluate the metabolic actions of SB, considering its physiologically relevant concentration. We evaluated the effect of SB on regulation of DPP-4 and its other metabolic actions, both in vitro (HepG2 cells and mouse mesangial cells) and in vivo (high fat diet [HFD]-induced obese mice). Ten-week HFD-induced obese C57BL/6J mice were subjected to SB treatment by adding SB to HFD which was maintained for an additional 16 weeks. In HepG2 cells, SB suppressed DPP-4 activity and expression at sub-molar concentrations, whereas it increased DPP-4 activity at a concentration of $1,000{\mu}M$. In HFD-induced obese mice, SB decreased blood glucose, serum levels of insulin and $IL-1{\beta}$, and DPP-4 activity, and suppressed the increase in body weight. On the contrary, various tissues including liver, kidney, and peripheral blood cells showed variable responses of DPP-4 to SB. Especially in the kidney, although DPP-4 activity was decreased by SB in HFD-induced obese mice, it caused an increase in mRNA expression of $TNF-{\alpha}$, IL-6, and $IL-1{\beta}$. The pro-inflammatory actions of SB in the kidney of HFD-induced obese mice were recapitulated by cultured mesangial cell experiments, in which SB stimulated the secretion of several cytokines from cells. Our results showed that SB has differential actions according to its treatment dose and the type of cells and tissues. Thus, further studies are required to evaluate its therapeutic relevance in metabolic diseases including diabetes and obesity.

• Animal cells and systems
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• 제15권3호
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• pp.227-233
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• 2011

The Nramp1/Slc11a1 locus encodes a proton-coupled divalent cation transporter, expressed in late endosomes/lysosomes of macrophages, that constitutes a component of the innate immune response to combat intracellular pathogens and it was shown to play an important role in regulating inherent immunity. The previously identified Z-DNA forming polymorphic repeat(GT)n in the promoter region of the human Nramp1 gene does act as a functional polymorphism influencing gene expression. Research has shown that INF-${\gamma}$, TNF-${\alpha}$, IL-$1{\beta}$ and bacteria LPS increase the level of Nramp1 expression. However, the molecular mechanism for Nramp1 gene regulation is unclear. In this research, bovine Nramp1 5'-flanking region (-1748~+769) was cloned and analyzed by bioinformatics. Then to find the core promoter and the cis-acting elements, deletion analysis of promoter was performed using a set of luciferase reporter gene constructs containing successive deletions of the bovine Nramp1 5'-flanking regions. Promoter activity analysis by the dual luciferase reporter assay system showed that the core promoter of Nramp1 was located at +58~-89 bp. Some positive regulatory elements are located at -89~-205 bp and -278~-1495 bp. And the repressor elements were in region -205~-278 bp, intron1 and -1495~-1748 bp. LPS-responsive regions were located at -1495~-1748 bp and -278~-205 bp. The present study provides an initial effort to explore the molecular mechanism of transcriptional activation of the bovine Nramp1 gene and should facilitate further studies to decode the complex regulatory process and for molecular breeding for disease resistance in bovines.

• IMMUNE NETWORK
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• 제2권1호
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• pp.25-34
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• 2002

Background: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial hyperplasia and joint destruction. The synovial fibroblasts express cell adhesion molecules and have a role in adhesive interation with inflammatory cells in synovial tissue. It has been suggested that hypoxic conditioins are thought to exist in arthritic joints, and several studies indicate that reactive oxygen species (ROS) produced in hypoxic condition can initiate events that lead to pro-adhesive changes via increased expression of adhesion molecules. So, this study wsa designed to examine whether antioxidant can inhibit hypoxia-induced expression of ICAM-1 in cultured human synovial fibroblasts. Methods: Synovial fibroblasts were isolated from synovial tissue in patients with RA and cultured at hypoxic condition. Antioxidant, PDTC (pyrrolidine dithiocarbamate) were pre-treated for an hour before the hypoxic culture and synovial fibroblasts were harvested at 0, 6, 12, 24, 48 hours time points. Cell surface ICAM-1 expression in synovial fibroblasts was examined by the flow cytometric analysis. To analyse the expression of ICAM-1 mRNA, reverse-transcriptase polymerase chain reaction (RT-PCR) was performed. The levels of cytokines in culture supernatants were measured by ELISA, and activation of NF-${\kappa}B$ was analysed by electrophoretic mobility shift assay. The adhesive reaction between synovial fibroblasts and lymphocytes was assayed by measurement of fluorescent intensity of BCECF-AM in lymphocytes. Results: Hypoxic stimuli up-regulated the ICAM-1 expression as well as the adhesive interaction of human synvial fibroblasts to lymphocytes in a time-dependent manner, and PDTC inhibited hpyoxia-induced ICAM-1 expression and cell-cell interaction. PDTC also inhibited the hypoxia-induced activation of intracellular transcription factor, NF-${\kappa}B$. PDTC decreased the amount of hypoxia-induced production of IL-$1{\beta}$ and TNF-${\alpha}$. Conclusion: These studies demonstrate that PDTC inhibit the hypoxia-induced expression of the adhesion molecule, ICAM-1 and activation of NF-${\kappa}B$ in cultured human synovial fibroblasts.

• Toxicological Research
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• 제34권2호
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• pp.103-110
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• 2018

Environmental stimuli can lead to the excessive accumulation of reactive oxygen species (ROS), which is one of the risk factors for premature skin aging. Here, we investigated the protective effects of $7-MEGA^{TM}$ 500 (50% palmitoleic acid, 7-MEGA) against oxidative stress-induced cellular damage and its underlying therapeutic mechanisms in the HaCaT human skin keratinocyte cell line (HaCaT cells). Our results showed that treatment with 7-MEGA prior to hydrogen peroxide ($H_2O_2$)-induced damage significantly increased the viability of HaCaT cells. 7-MEGA effectively attenuated generation of $H_2O_2$-induced reactive oxygen species (ROS), and inhibited $H_2O_2$-induced inflammatory factors, such as prostaglandin $E_2$ ($PGE_2$), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), and $interleukin-1{\beta}$ ($IL-1{\beta}$). In addition, cells treated with 7-MEGA exhibited significantly decreased expression of matrix metalloproteinase-1 (MMP-1) and increased expression of procollagen type 1 (PCOL1) and Elastin against oxidative stress by $H_2O_2$. Interestingly, these protective activities of 7-MEGA were similar in scope and of a higher magnitude than those seen with 98.5% palmitoleic acid (PA) obtained from Sigma when given at the same concentration (100 nL/mL). According to our data, 7-MEGA is able to protect HaCaT cells from $H_2O_2$-induced damage through inhibiting cellular oxidative stress and inflammation. Moreover, 7-MEGA may affect skin elasticity maintenance and improve skin wrinkles. These findings indicate that 7-MEGA may be useful as a food supplement for skin health.

• 동의생리병리학회지
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• 제20권4호
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• pp.844-852
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• 2006

This study was to evaluate the effect of Yeongyupaedog-san (YGPDS) on mouse Thl and Th2 cells' differentiation and ovalbumin (OVA)-induced allergic inflammation. The proliferation of mouse CD4 T cells and the secretion of Th1/Th2 cytokines under the influence of YGPDS extract were measured as well as the amount of ${\beta}-hexosaminidase$ in RBL-2H3 cells and the levels of $TNF-{\alpha}$ and 1L-6 secretion in Raw264.7 cells. BALB/c mice were orally administered with YGPDS extract and simultaneously inoculated with OVA to induce allergic reaction and measure the level of total IgE, OVA-specific IgE and the production of IFN- g, IL-4, IL-5 by the spleen cells. When mouse CD4 T cell were stimulated with anti-CD3 and anti-CD28 for 48 hours in various concentrations of YGPDS extract, it increased proliferation of CD4 cells by 11% in $100\;{\mu}g/^{ml}$ concentration but it showed an inhibition by 37% at $200\;{\mu}g/^{ml}$ CD4 T cells under Th1/Th2 polarizing conditions for 3 days with YGPDS resulted in mild decrease of IFN- g in Thl cells and significant decrease of IL-4 in Th2 cells at $500\;{\mu}g/^{ml}\;and\;100\;{\mu}g/^{ml}$ by 18% and 21%, respectively. YGPDS extract had a dose-dependent inhibitory effect on antigen-induced release of ${\beta}-hexosaminidase$ in RBL-2H3 cells. Treatment of YGPDS extract on LPS stimulated Raw 264.7 cells showed dose-dependent decrease in TNF-n production. Oral administration of YGPDS extract on OVA-induced allergic mice showed an inhibitory effect on the levels of total serum IgE and OVA-specific IgE by 25% and 34% , respectively. Culture of spleen cells with OVA resulted in significant increase of IFN- g by 44% and significant decrease of IL-4 and IL-5 by 56%, and 24%, respectively. The results show that YGPDS does not strongly induce mouse T cells to transform into Thl or Th2 but it has an anti-allergic effect in vitro, and that it also corrects the unbalance between the reactions of Th cells in allergic diseases.

• Biomolecules & Therapeutics
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• 제25권3호
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• pp.279-287
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• 2017

The chemical property of cinnamaldehyde is unstable in vivo, although early experiments have shown its obvious therapeutic effects on viral myocarditis (VMC). To overcome this problem, we used cinnamaldehyde as a leading compound to synthesize derivatives. Five derivatives of cinnamaldehyde were synthesized: 4-methylcinnamaldehyde (1), 4-chlorocinnamaldehyde (2), 4-methoxycinnamaldehyde (3), ${\alpha}$-bromo-4-methylcinnamaldehyde (4), and ${\alpha}$-bromo-4-chlorocinnamaldehyde (5). Neonatal rat cardiomyocytes and HeLa cells infected by coxsackievirus B3 (CVB3) were used to evaluate their antiviral and cytotoxic effects. In vivo BALB/c mice were infected with CVB3 for establishing VMC models. Among the derivatives, compound 4 and 5 inhibited the CVB3 in HeLa cells with the half-maximal inhibitory concentrations values of $11.38{\pm}2.22{\mu}M$ and $2.12{\pm}0.37{\mu}M$, respectively. The 50% toxic concentrations of compound 4 and 5-treated cells were 39-fold and 87-fold higher than in the cinnamaldehyde group. Compound 4 and 5 effectively reduced the viral titers and cardiac pathological changes in a dose-dependent manner. In addition, compound 4 and 5 significantly inhibited the secretion, mRNA and protein expressions of inflammatory cytokines TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in CVB3-infected cardiomyocytes, indicating that brominated cinnamaldehyde not only improved the anti-vital activities for VMC, but also had potent anti-inflammatory effects in cardiomyocytes induced by CVB3.

• International Journal of Oral Biology
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• 제42권1호
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• pp.17-23
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• 2017

Background: Periodontitis is generally a chronic disorder characterized by the breakdown of tooth-supporting tissues. P. gingivalis, a Gram-negative anaerobic rod, is one of the major pathogens associated with periodontitis. Frequently, P. gingivalis infection leads to cell death. However, the correlation between P. gingivalis-induced cell death and periodontal inflammation remains to be elucidated. Among cell deaths, the death of immune cells appears to play a significant role in inflammatory response. Thus, the aim of this study was to examine P. gingivalis-induced cell death, focusing on autophagy and apoptosis in THP-1 cells. Methods: Human acute monocytic leukemia cell line (THP-1) was used for all experiments. Autophagy induced by P. gingivalis in THP-1 cells was examined by Cyto ID staining. Intracellular autophagic vacuoles were observed by fluorescence microscopy using staining Acridine orange (AO); and 3-methyladenine (3-MA) was used to inhibit autophagy. Total cell death was measured by LDH assay. Cytokine production was measured by an ELISA method. Results: P. gingivalis induced autophagy in an MOI-dependent manner in THP-1 cells, but 3-MA treatment decreased autophagy and increased the apoptotic blebs. P. gingivalis infection did not increase apoptosis compared to the control cells, whereas inhibition of autophagy by 3-MA significantly increased apoptosis in P. gingivalis-infected THP-1 cells. Inhibition of autophagy by 3-MA also increased total cell deaths and inflammatory cytokine production, including $IL-1{\beta}$ and $TNF-{\alpha}$. Conclusion: P. gingivalis induced autophagy in THP-1 cells, but the inhibition of autophagy by 3-MA stimulated apoptosis, leading to increased cell deaths and pro-inflammatory cytokines production. Hence, the modulation of cell deaths may provide a mechanism to fight against invading microorganisms in host cells and could be a promising way to control inflammation.

• 생명과학회지
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• 제24권11호
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• pp.1157-1167
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• 2014

잡곡은 대사성 질환의 예방과 치료에 우수한 기능성 식품소재로 인식되고 있다. 국내산 잡곡들(기장, 황금찰수수, 노랑차조, 율무 및 식용피)을 대상으로 항염증 활성을 비교조사하고자, 이들 잡곡의 80% 에탄올 추출물을 이용하여 마우스 대식세포주 RAW264.7을 지질다당류(LPS)로 자극할 때 생성되는 염증 매개인자인 산화질소(NO)의 생성에 미치는 저해능을 조사하였다. 그 결과, 식용피(Echinochloa crus-galli var. frumentacea) 유래의 에탄올 추출물이 LPS에 의해 유도되는 NO의 생성을 저해하는 항염증 활성에 있어서 가장 우수함을 확인하였다. 식용피의 에탄올 추출물로부터 항염증 관련 성분을 규명하기 위해, 80%에탄올 추출물을 물에 녹인 후 n-hexane, methylene chloride (MC), ethyl acetate (EtOAc), n-butanol (BuOH)과 물 분획들로 단계별 분획하고, 각 분획의 항염증 활성을 조사하였다. RAW264.7 세포에 MC 분획($100{\mu}g/ml$)을 전처리하였을 때, LPS처리에 의해 유도되는 iNOS, COX-2, 그리고 친염증성 사이토카인들(IL-$1{\beta}$, IL-6, 및 TNF-${\alpha}$)의 발현이 현저하게 저해되는 것으로 나타났다. 또한, LPS처리에 의해 유도되는 p38MAPK, JNK 및 ERK의 활성화의 경우도 MC 분획에 의해 농도-의존적으로 저해되는 것으로 나타났다. HPLC분석을 통해, 식용피 유래 MC 분획의 항염증 활성은 식용피의 MC 분획 속의 주요성분인 kaempferol과 biochanin A에 기인함을 확인하였다. 이상의 연구 결과는 식용피 및 식용피의 MC 분획이 염증성 질환과 이와 관련된 대사성 질환의 예방과 예후개선에 유리한 기능성 식품소재 개발에 활용될 수 있음을 시사한다

• 한국식품영양과학회지
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• 제44권11호
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• pp.1612-1620
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• 2015

양파 껍질 에탄올 추출물(OPEE)의 in vitro 및 in vivo에서 항염증 효과를 알아보기 위해 lipopolysaccharide(LPS)로 활성화된 RAW 264.7 세포로부터 분비되는 염증 관련 매개인자들의 발현량과 ICR 마우스모델을 이용한 귀 부종 및 조직 분석을 진행하였다. NO 및 염증성 cytokine(IL-6, $IL-1{\beta}$ 및 $TNF-{\alpha}$)의 분비량이 OPEE에 의해 농도 의존적으로 감소함을 확인하였다. iNOS 및 COX-2의 발현량은 LPS에 의해 증가하였으나, OPEE 처리에 의해 농도 의존적으로 발현량이 감소하였다. 전사인자인 $NF-{\kappa}B$의 활성과 인산화효소인 MAPKs(p-38, JNK 및 ERK)에서도 LPS에 의해 증가된 발현량이 OPEE에 의해 억제됨을 확인하였다. 따라서 NO 및 염증성 cytokine의 분비량 감소는 $NF-{\kappa}B$와 MAPKs의 활성 저해에 의한 것임을 확인하였다. In vivo 실험에서는 croton oil로 귀 부종을 유발한 마우스에 OPEE를 처리하였을 때 prednisolone 처리구와 유사한 수준으로 귀 부종이 억제되었으며, 귀 조직 관찰에서도 경피 및 진피의 두께와 진피로 침윤된 mast cell의 수도 억제됨을 확인하였다. 이상의 결과 OPEE는 항염증 소재로서 염증을 예방하거나 치료하는 데 이용 가능성이 높음을 제시한다.