• 생약학회지
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• 제35권1호통권136호
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• pp.92-97
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• 2004

Effect of AGI $({\alpha}-Glucosidase\;Inhibitor)-1120$, pine (Pinus densiflora) bark extract and Chaga mushroom (Inonotus obliquus) - and Chaga mushroom mycelium extracts on cellular $NF-{\kappa}B$ activation in malignant human keratinocytes (SCC-13) were evaluated to elucidate the possible correlation of $NF-{\kappa}B$ with antioxidant activity. The antioxidant activities of these natural products were examined in three different evaluation methods, i.e., lipid peroxidation value (POV) evaluation test, and 1,1diphenyl-2-picrylhydrazyl radical (DPPH) and nitric oxide (NO) scavenging test. In a cell-based $NF-{\kappa}B$ monitoring assay systern, all samples revealed the downregulatory profiles on the cellular $NF-{\kappa}B$ activity. AGI -1120 (1, 2 mg) and Chaga mushroom extract (0.05, 0.1 mg) downregulated the $NF-{\kappa}B$ activity in a dose-dependent manner. Chaga mushroom mycelium extract (5 mg) significantly inhibited the $NF-{\kappa}B$ activity (p<0.05). Although AGI-1120 and Chaga mushroom mycelium extract exhibited no antioxidant activities evaluated in pay, Chaga mushroom extract showed antioxidant in a dose-dependent manner at concentrations of $0.05{\sim}1$ mg. While AGI-1120 and Chaga mushroom extract possessed a relatively potential DPPH radical scavenging activity, the NO scavenging activity of Chaga mushroom extract $(SC_{50}:47\;{mu}g)$ was higher than the known antioxidant, vitamin C $(SC_{50}:77\;{mu}g)$. These results suggest that AGI-1120 and Chaga mushroom- and Chaga mushroom mycelium extracts may serve as an useful radical scavenging antioxidant agents with $NF-{\kappa}B$ inhibitory effect in human skin.

• 한국수산과학회지
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• 제47권6호
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• pp.757-764
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• 2014

Many researchers have studied algae as a source of material having potential biological activities, not least because many marine algae extracts have strong antioxidant properties. In this study, we investigated the anti-inflammatory effects of Pyropia yezoensis extract (PYE) on RAW 264.7 cells by measuring nitric oxide (NO), reactive oxygen species (ROS), superoxide dismutase (SOD), catalase activity, inducible NOS (iNOS), cyclooxygenase-2 (COX-2), nuclear factor-kappa B (NF-${\kappa}B$), interleukin-$1{\beta}$ (1L-$1{\beta}$), and tumor necrosis factor-alpha (TNF-${\alpha}$). PYE decreased the production of intracellular ROS dose-dependently and increased SOD and catalase activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. PYE significantly suppressed the production of NO and reduced the expression of iNOS, COX-2, and NF-${\kappa}B$. PYE treatment also inhibited the production of IL-$1{\beta}$ and TNF-${\alpha}$ significantly and reduced the phosphorylation of Akt and MAPK significantly in LPS-stimulated RAW 264.7 cells. These results suggest that PYE has potential anti-oxidant and anti-inflammatory activities.

• BMB Reports
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• 제47권6호
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• pp.318-323
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• 2014

We isolated the phenolic glucoside salicortin from a Populus euramericana bark extract, and examined its ability to suppress inflammatory responses as well as the molecular mechanisms underlying these abilities, using lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Salicortin inhibited iNOS expression and the subsequent production of NO in a dose-dependent manner in the LPS-stimulated RAW 264.7 cells. Salicortin significantly suppressed LPS-induced signal cascades of NF-${\kappa}B$ activation, such as IKK activation, $I{\kappa}B{\alpha}$ phosphorylation and p65 phosphorylation in RAW 264.7 cells. In addition, salicortin inhibited the LPS-induced activation of JNK, but not ERK or p38 MAPK. Furthermore, salicortin significantly inhibited production of pro-inflammatory cytokines, such as TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in the LPS-stimulated RAW 264.7 cells. These findings suggest that salicortin may show its anti-inflammatory activity by suppressing the LPS-induced expression of pro-inflammatory mediators through inhibition of NF-${\kappa}B$ and JNK MAPK signaling cascades in macrophages.

• Advances in Traditional Medicine
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• 제10권3호
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• pp.155-164
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• 2010

Lycii fructus is the fruit of Lycium chinense Miller and is part of the Solanaceae family. Lycii fructus produces various effects such as hypotensive, hypoglycemic, anti-pyretic, and anti-stress activities. Lycii fructus is known to contain betaine, carotene, nicotinic acid, zeaxanthin, and cerebroside. In the present study, the effects of Lycii fructus aqueous extract on lipopolysaccharide (LPS)-induced inflammation in murine raw 264.7 macrophage cells were investigated. In this study we utilized the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcriptionpolymerase chain reaction (RT-PCR), Western blotting, and nitric oxide (NO) detection. Lycii fructus aqueous extract suppressed NO production by inhibiting the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-$\alpha$) mRNA and iNOS protein in murine raw 264.7 macrophage cells. Also, Lycii fructus aqueous extract suppressed the activation of nuclear factor-kappa B (NF-${\kappa}B$) in the nucleus. These results demonstrated that Lycii fructus aqueous extract causes an anti-inflammatory effect that was likely produced by the suppression of iNOS expression through the down-regulation of NF-$\hat{e}B$ binding activity.

• Journal of Applied Biological Chemistry
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• 제52권1호
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• pp.21-27
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• 2009

인삼은 가공처리방법에 따라 홍삼과 발효홍삼으로 구분된다. 또한, 이들의 가공처리방법에 따라 인삼의 효능을 나타내는 사포닌의 함량이 차이가 있다. 따라서, 본 연구에서 인삼, 홍삼 및 발효홍삼이 대식세포에서 LPS에 의한 염증에 대한 항염효과 및 그 기전을 규명하고자 하였다. 마우스의 대식세포인 RAW264.7 세포에서 LPS에 의해 유도되는 염증관련인자인 NO 및 COX-2의 발현 및 TNF-$\alpha$, INF-$\gamma$ 그리고 NF-${\kappa}B$의 활성을 인삼, 홍삼 그리고 발효홍삼에 의한 항염효과 차이를 비교 하였다. 그 결과, 인삼 및 홍삼 그리고 발효홍삼 모두에서 LPS에 의한 NO의 생성을 억제시키는 것을 확인하였으며, TNF-$\alpha$ 및 INF-$\gamma$의 생성 또한 억제시키는 것을 알 수 있었다. 또한, 인삼 및 홍삼 그리고 발효홍삼 모두 COX-2의 발현 및 LPS에 의한 $I{\kappa}B$의 인산화를 억제시킴으로써 NF-${\kappa}B$의 활성을 억제시키는 것임을 알 수 있었다. 홍삼이 인삼과 발효홍삼에 비하여 NO의 생성을 더 효과적으로 억제시키는 것은 각각의 제조과정에서 나타나는 인삼사포닌의 조성의 차이에 따른 것으로 추정된다. 본 연구는 단순한 동물세포 수준에서의 비교 차이이며 좀 더 정확한 기전의 규명을 위해서는 향후 동물실험을 통한 비교 실험이 수행되어야 할 것이다.

• 대한본초학회지
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• 제34권1호
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• pp.51-57
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• 2019

Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

• 대한본초학회지
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• 제30권1호
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• pp.85-93
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• 2015

Objectives : Bilobalide (BIL) is a predominant sesquiterpene trilactone constituent that accounts for a partial portion of the standardized Ginkgonis Folium extract, which has been widely used to treat a variety of neurological disorders involving cerebral ischemia and neurodegeneration. In this study, it was tested whether BIL exhibits anti-inflammatory activities on inflammation response, or not. Methods : To elucidate the molecular mechanisms of BIL on pharmacological and biochemical actions in inflammation, we examined the effect of BIL on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophages. The investigation was focused on how BIL affect on inflammation-related mediators including various signals such as nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), inducible NO synthase(iNOS), cyclooxygenase-2(COX-2), interleukin-6(IL-6), tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), mitogen-activated protein kinases(MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$) in LPS-stimulated RAW 264.7 cells. Results : We found that BIL inhibited LPS-induced NO, $PGE_2$, IL-6 and $TNF-{\alpha}$ productions as well as the expressions of iNOS and COX-2. Furthermore, BIL suppressed the LPS-induced phosphorylation for MAPK activation. Conclusions : These results suggest that BIL has inhibitory effects on LPS-induced $PGE_2$, NO, IL-6 and $TNF-{\alpha}$ production, as well as the expressions of iNOS and COX-2 in the murine macrophage. It seems that these inhibitory effects occur by blocking the phosphorylation of MAPKs for activation. Then, BIL suppressed the activation of nuclear factor $NF-{\kappa}B$ in nucleus. These observations suggest that BIL has anti-inflammatory effect by inhibiting.

• BMB Reports
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• 제42권6호
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• pp.380-386
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• 2009

In neurodegenerative diseases, such as Alzheimer' and Parkinson', microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds. Prothrombin and kringle-2 increase levels of NO and the mRNA expression of iNOS, IL-1$\beta$, and TNF-$\alpha$ in microglial cells. In contrast, the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1$\beta$, TNF-$\alpha$, and IL-6 in LPS-activated EOC2 microglia. In this study, we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia. Treatment with 20-100 ${\mu}M$ of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation. NO production induced by MAPKs and NF-$\kappa$B was similarly reduced by inhibitors of ERK (PD98059), p38 (SB203580), NF-$\kappa$B (N-acetylcysteine), and NSA9. These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2.

• Journal of Acupuncture Research
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• 제23권6호
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• pp.117-132
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• 2006

Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

• The Korean Journal of Physiology and Pharmacology
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• 제18권4호
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• pp.313-320
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• 2014

The study was conducted to investigate the role of vitamin E in the high altitude hypoxia-induced damage to the intestinal barrier in rats. Sprague-Dawley rats were divided into control (Control), high altitude hypoxia (HH), and high altitude hypoxia + vitamin E (250 mg/kg $BW^*d$) (HV) groups. After the third day, the HH and HV groups were placed in a hypobaric chamber at a stimulated elevation of 7000 m for 5 days. The rats in the HV group were given vitamin E by gavage daily for 8 days. The other rats were given equal volume saline. The results showed that high altitude hypoxia caused the enlargement of heart, liver, lung and kidney, and intestinal villi damage. Supplementation with vitamin E significantly alleviated hypoxia-caused damage to the main organs including intestine, increased the serum superoxide dismutase (SOD) (p< 0.05), diamino oxidase (DAO) (p< 0.01) levels, and decreased the serum levels of interleukin-2 (IL-2) (p< 0.01), interleukin-4 (IL-4) (p<0.001), interferon-gamma ($IFN-{\gamma}$) (p<0.01) and malondialdehyde (MDA) (p<0.001), and decreased the serum erythropoietin (EPO) activity (p<0.05). Administration of vitamin E significantly increased the S-IgA (p<0.001) in ileum and significantly improved the expression levels of occludin and $I{\kappa}B{\alpha}$, and decreased the expression levels of hypoxia-inducible factor 1 alpha and 2 alpha ($HIF-1{\alpha}$ and $HIF-2{\alpha}$), Toll-like receptors (TLR4), P-$I{\kappa}B{\alpha}$ and nuclear factor-${\kappa}B$ p65(NF-${\kappa}B$ P65) in ileum compared to the HH group. This study suggested that vitamin E protectis from intestinal injury caused by high altitude hypoxia environment. These effects may be related to the HIF and TLR4/NF-${\kappa}B$ signaling pathway.