• 한국수산과학회지
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• 제46권4호
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• pp.384-392
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• 2013

An ethanolic extract of Undaria pinnatifida was fractionated using several solvents. Of the fractions, the ethyl acetate fraction had the greatest inhibitory effect on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophage cells. Using this fraction (U. pinnatifida ethyl acetate extract, UPE), we investigated the molecular mechanism underlying its inhibitory effect on LPS-stimulated RAW 264.7 cells. Pretreatment of the cells with up to $100{\mu}g/mL$ UPE significantly inhibited NO production and inducible nitric oxide synthase (iNOS) expression, in a dose-dependent manner. Similarly, UPE treatment markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1, IL-6 and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), while it strongly suppressed the nuclear translocation of nuclear factor-kappa B (NF-${\kappa}B$) by preventing proteolytic degradation of inhibitor of nuclear factor ${\kappa}B$ $(I{\kappa}B)-{\alpha}$. Moreover, UPE treatment significantly reduced the phosphorylation of phosphatidylinositol 3-kinase (PI3K)/Akt and mitogen-activated protein kinase (MAPK) in LPS-stimulated cells. These results indicate that UPE contains anti-inflammatory compounds and suggest that it might be used as a functional food material that assists in prevention of inflammatory diseases.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.101-101
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• 2003

Prostaglandins (PGs) and nitric oxide (NO) produced by inducible cyclooygenase (COX-2) and nitric oxide synthase (iNOS), respectively, have been implicated as important mediators in the process of inflammation and carcinogenesis. On this line, the potential COX-2 or iNOS inhibitors have been considered as anti-inflammatory and cancer chemopreventive agents. In our continuing efforts of searching for novel cancer chemopreventive agents from natural products, we isolated natural sesquiterpenoids as potential COX-2 and iNOS inhibitors in cultured lipopolysaccharide (LPS)-activated mouse macrophage RAW 264.7 cells. Alantolactone, a natural eudesmane-type sesquiterpenoid, exhibited a potent inhibition of COX-2 (IC50 = 0.4 $\mu\textrm{g}$/$m\ell$) and iNOS activity (IC50 = 0.08 $\mu\textrm{g}$/$m\ell$) in the assay system determined by PGE2 and NO accumulation, respectively. The inhibitory potential of alantolactone on the PGE2 and NO production was well coincided with the suppression of COX-2 and iNOS protein and mRNA expression in LPS-induced macrophages. Furthermore, alantolactone inhibited NF-kB but not AP-l binding activity on nuclear extracts evoked by LPS-stimulated macrophage cells, suggesting the possible involvement of NF-kB in the regulation of COX-2 and iNOS expression. In further study with COX-2-expressing human colon HT-29 cells, alantolactone inhibited the cell proliferation, down-regulated COX-2, and inhibited the ERK phosphorylation in the early time. These results suggest that a natural sesquiterpenoid alantolactone might be a potential lead candidate for further developing COX-2 or iNOS inhibitor possessing cancer chemopreventive or anti-inflammatory activity

• Food Science and Biotechnology
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• 제15권6호
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• pp.975-979
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• 2006

The extracts of Oenanthe javanica were evaluated for their effects on the expression of cyclooxygenase-2 (COX-2), which is mediated by the translocation of the p65 subunit into the nucleus. Fractions of ethyl acetate and chloroform from 80% ethanol extracts of O. javanica exhibited inhibitory effects on the secretion of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) from lipopolysaccharide (LPS)-stimulated peritoneal macrophages; however, the aqueous- and hexane-fractions showed no significant effect. The ethyl acetate- and chloroform-fractions also reduced the COX-2 enzyme levels after 24-hr treatment. RT-PCR showed that the mRNA levels of COX-2 decreased following treatment with these fractions, suggesting that COX-2 expression is transcriptionally regulated by these extracts. We examined the effects of the chloroform- and ethyl acetate-fractions on the cytosolic activation of nuclear factor-${\kappa}B$ ($NF-{\kappa}B$, p65 subunit) and on the degradation of inhibitor-${\kappa}B{\alpha}$ ($I-{\kappa}B{\alpha}$) in order to determine the mechanism of COX-2 regulation. The LPS-stimulated activation of the p65 subunit was significantly blocked upon the addition of $50\;{\mu}g/mL$ of these fractions, and the cytosolic $I-{\kappa}B{\alpha}$ degradation process was simultaneously inhibited. These findings suggest that the inhibition of COX-2 expression by the ethyl acetate-and chloroform-fractions may result from the inhibition of p65 translocation by blocking the degradation of $I-{\kappa}B{\alpha}$; this may be the mechanistic basis for the anti-inflammatory effects of O. javanica.

• Food Science and Biotechnology
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• 제18권5호
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• pp.1063-1070
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• 2009

Dangyuja (Citrus grandis Osbeck) is a native plant growing only on Jeju Island in Korea. In this study, antiinflammatory effect of dangyuja leaves on a murine macrophage cell line was investigated. RAW 264.7 murine macrophage cells were stimulated with lipopolysaccharide (LPS, $1{\mu}g/mL$) to induce expression of pro-inflammatory markers [interleukin (IL)-6 and inducible nitric oxide synthase (iNOS)]. The crude extract (80% MeOH Ex.) and solvent fractions (hexane, $CHCl_3$, EtOAc, BuOH, and $H_2O$ Ex.) were obtained from dangyuja leaves. The $CHCl_3$ fraction inhibited the nitric oxide (NO) and IL-6 production in a dose-dependent manner. Also, the $CHCl_3$ fraction inhibited mRNA expression and protein levels of iNOS in a dose-dependent manner. Furthermore, the $CHCl_3$ fraction inhibited LPS-induced nuclear factor (NF)-${\kappa}B$ activation and phosphorylation of mitogen-activated protein kinases (MAPKs: ERK, JNK, and p38). These results suggest that dangyuja leaves may inhibit LPS-induced production of inflammatory markers by blocking NF-${\kappa}B$ and MAPKs signaling in RAW 264.7 cells.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2010년도 정기총회 및 추계학술발표회
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• pp.7-7
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• 2010

전 세계적으로 폭발적으로 증가하고 있는 비만은 만병의 근원이다. 동시에 대사성질환의 발병을 증가시키는 결정적인 역할을 하고 있다. 이러한 상황을 극복할 수 있는 안전하고 효능이 우수한 약의 개발이 매우 시급하다. 시판되는 약들은 예견할 수 없는 지방변으로 실제생활을 매우 불편하게 하는 부작용과 우울증 및 자살충동 등의 심각한 부작용을 유발시키고 있다. 특히 생명을 위협하는 약은 시판이 금지되었다. 200여종의 한약재들을 In vitro screening (pancreatic lipase inhibition, PDE inhibition, c-AMP activity), ex vivo screening (lipolytic action on fat pad), short term animal screening(혈중 TG 함량 분석)을 토대로 long term animal model에서비만 예방 및 치료 효능을 검증하기 위하여 호장근 부탄올 분획을 선정하였다. 고지방 사료로 비만을 유도한 rat (Diet induced obesty (DIO) rat)에서 비만 치료 효능이, 고지방 사료로 비만을 유도한 ICR-mice에서 비만 예방 효능이 우수함을 입증하였다 (체중감소, 지방세포의 크기억제, 지방간 예방/치료(간무게, TG함량, 간 색상, 고지혈증 증상억제), 혈중 TNF-a, IL-6, leptin, adponectin 등, 간 조직에서의 pAMPK, SOCS, NF-${\kappa}B$ DNA biding activity, ACC level, FAS expression, CPT-1 activity의 정상화). 호장근 부탄올 분획의 이러한 효능은 AMPK 작용과 CPT-1 작용을 활성화하고 동시에 지방산 합성 억제와 지방산 산화를 촉진함으로 인함임을 규명하였다. 동시에 비만으로 인한 pancreatic beta cell의 파괴를 예방함으로 인슐린 내성을 예방(치료)함을 입증하였다. 이는 AMPK 활성화와 SOCS-3 단백질 억제와 NF-${\kappa}B$-DNA 결합 억제로 인함임을 증명하였다. 3T3-L1 지방 세포주에서 lipogenesis 예방(치료) 및 lipolytic effect에 관여하는 인자들의 변화를 확인하였다. 이는 Multi-compounds-multi-targets에 의한 시너지 효과임을 알 수 있었다.

• 대한한의학회지
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• 제26권1호
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• pp.46-58
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• 2005

Objective : This study on Sipimikwanjung-tang was undertaken to evaluate its antioxidant capacities and antiperoxidation activities in rat liver tissues. Sipimikwanjung-tang which has been one of the prescriptions in sasang constitutional medicine is usually applied for the therapy of various liver diseases. It is elucidated that Sipimikwanjung-tang has antioxidants on liver tissue of rat and the cytotoxic effects on human hepatoblastoma Hep G2 cells. Methods: Sipimikwanjung-tang extract in antioxidant effects of Hep G2 cells is evaluated by MTT assay, DAPI staining, DNA fragmentation assays and FACS can analysis. Results: Sipimikwanjung-tang induced apoptosis in Hep G2 cells, and induced G1 and G2M arrest of the cell cycle as well as a significant increase in PARP and caspase-3 activity. It induced an increase in $H_2O_2$ generation and the subsequent $NF-{\kappa}B$ activation and also induced cell apoptosis through the caspase-3-dependent pathways in the low concentration of Sipimikwanjung-tang extracts. However, the high dose of Sipimikwanjung-tang extract in Hep G2 cells inhibited $TGF-{\beta}l-induced$ apoptosis via increase in cellular $H_2O_2$, formation and $NF-{\kappa}B$ activation in human hepatoblastoma Hep G2 cells. Conclusion: From this study, the possibility that Sipimikwanjung-tang extracts apply to antioxidant and apoptotic treatment of disease is revealed.

• 대한한의학회지
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• 제36권1호
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• pp.9-21
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• 2015

Objectives: Realgar has been frequently used for skin disorders in history of herbal medicine. However, the efficacy of realgar has not been examined in atopic dermatitis(AD). In this study, the effects of realgar on AD were investigated, especially on pruritus and inflammation. Methods: AD lesions were induced in the shaved backs of BALB/c mice through repeated application of DNCB. The mice were treated for 11 days with 1% realgar ($100{\mu}L/day$). Histological changes in skin thickness were observed. The anti-pruritic effects of realgar were evaluated by the change in numbers of scratching behavior of mice and expression of substance P. The expressions of cytokines IL-4 and IL-6 were measured. Also, anti-inflammatory effects of realgar were examined on expressions of NF-${\kappa}B$, phospho-$I{\kappa}B{\alpha}$ and mitogen-activated protein kinases (MAPKs). Results: Realgar decreased skin thickness (both dermal and epidermal) 38% and 17% respectively, compared to positive control, DNCB group. The scratching behavior of mice was reduced by 42% and expression of substance P was significantly less. Cytokines IL-4 and IL-6 were significantly reduced by 52.6% and 77.6%, respectively. The expressions of NF-${\kappa}B$, phospho-$I{\kappa}B{\alpha}$ and MAPKs (phospho-ERK1/2, -p38 and -JNK) were significantly suppressed with marked effects on phospho-ERK1/2. Conclusions: The collective results suggest that realgar shows anti-pruritic and anti-inflammatory effects on AD. And realgar might be a potential therapeutic candidate for treatment of atopic dermatitis.

• 대한한방부인과학회지
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• 제24권2호
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• pp.13-32
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• 2011

Objectives: This study was performed to elucidate the effects of Danchisoyo-san (DS) on cell damage and gene expression in UVB-exposed dermal fibroblast. Methods: To demonstrate the inhibitory effects of DS on aging of the skin, we used human dermal fibroblast(F6) and UVB light(30 mJ/$cm^2$) was used to damage to dermal fibroblast. We measured the nitrite production, LDH release, and gene expression in UVB-irradiated dermal fibroblast to elucidate the actionmechanism of DS. Also, we evaluated the amount of increased PICP, TIMP-1 in dermal fibroblast. PICP, TIMP-1 concentration was measured using EIA kit, and gene expression (MMP-1, procollagen, c-fos, c-jun, NF-kB, Bcl-2, Bcl-xL, iNOS) were determined using real-time PCR. Results: 1. DS inhibited LDH-release, nitrite production in UVB-irradiated dermal fibroblast. 2. DS suppressed the gene expression of MMP-1 in UVB-irradiated dermal fibroblast. 3. DS increased the gene expression of procollagen in UVB-iradiated dermal fibroblast. 4. DS suppressed the gene expression of c-jun, c-fos, NF-kB, iNOS in UVBirradiated dermal fibroblast. 5. DS increased the gene expression of Bcl-2 in UVB-iradiated dermal fibroblast. 6. DS increased the cell proliferation of dermal fibroblast. Conclusions: From the results, we concluded DS increases the cell proliferation and collagen synthesis in dermal fibroblast. So we suggest that DS has the antiwrinkle effects.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1193-1203
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• 2019

We investigated the protective effects of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey against hydrogen peroxide ($H_2O_2$)-induced apoptosis in Vero cells. BDB exhibited scavenging activity for DPPH, hydroxyl, and alkyl radicals. BDB also inhibited $H_2O_2$-induced lipid peroxidation, cell death, and apoptosis in Vero cells by inhibiting the production of ROS. To evaluate the molecular mechanisms of apoptosis inhibition, the expression of Bax/Bcl-xL and $NF-{\kappa}B$ was assessed by western blot assay. BDB significantly suppressed the cleavage of caspase-9 and PARP and reduced Bax levels in $H_2O_2$-induced Vero cells. Besides, BDB suppressed the phosphorylation of $NF-{\kappa}$B and the translocation of p65 in $H_2O_2$-induced cells. Furthermore, we evaluated the effect of BDB on ROS production, cell death, and lipid peroxidation in an $H_2O_2$-stimulated zebrafish embryo model. Taken together, these results indicated that ROS generation and cell death were significantly inhibited by BDB in zebrafish embryos, thereby proving that BDB exerts excellent antioxidant activity in vitro and in vivo.

• 한국수산과학회지
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• 제54권2호
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• pp.152-161
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• 2021

Eisenia bicyclis is known to have secondary metabolites exhibiting various biological activities. In a preliminary study, the n-hexane fraction obtained from the ethanolic extract of E. bicyclis showed higher anti-inflammatory activity than the ethyl acetate and butyl alcohol fractions based on the inhibition of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 cells. Using this fraction (E. bicyclis hexane fraction, EHF), we investigated the molecular mechanisms underlying its anti-inflammatory effect in LPS-stimulated RAW 264.7 cells. Pretreatment of the cells with up to 50 ㎍/mL EHF significantly inhibited NO and prostaglandin E₂ production as well as their responsible enzyme proteins and mRNAs, in a dose-dependent manner (P<0.05). Similarly, EHF markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α as well as their mRNA levels. Nuclear translocation of nuclear factor-kappa B (NF-κB) was strongly suppressed by EHF treatment. EHF significantly reduced the phosphorylation of mitogen-activated protein kinases and phosphatidylinositol 3-kinase/Akt in LPS-stimulated cells. Moreover, EHF reduced ear edema in phorbol myristate acetate (PMA)-induced mice. These results indicate that EHF contains potent anti-inflammatory compounds, which may be used as a dietary supplement for the prevention of inflammatory diseases.