• Journal of Korean Medicine Rehabilitation
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• 제24권4호
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• pp.15-28
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• 2014

Objectives This study was carried out to find out the Antioxidant and Anti-inflammatory Effects of Components of Mahwangbujaseshin-tang in LPS-Stimulated RAW264.7 Macrophages. Methods There are 5 experimental groups. ; normal, control, EH (Ephedrae Herba), ALRP (Aconiti Lateralis Radix Preparata) and AR (Asiasari Radix). The extract of EH, ALRP and AR ($100{\mu}g/ml$) was added to each group. We examined cytotoxicity, total phenolic contents, DPPH and ABTS free radical scavenging activity, Intracellular ROS (reactive oxygen species) production, NO (Total Nitric oxide), iNOS (inducible nitric oxide synthase), PGE2 (prostaglandin E2), COX-2 (cyclooxygenase-2), $IL-1{\beta}$ ($interleukin-1{\beta}$), IL-6 (interleukin-6), $TNF-{\alpha}$ (tumor necrosis factor-${\alpha}$), MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinase-1) and HO-1 (heme oxygenase-1) expression level. Results 1. Total phenolic contents of EH were in the highest level. 2. DPPH and ABTS free radical scavenging activity of EH was in the highest level. 3. ROS production was significantly decreased in AR. 4. NO production was significantly decreased in EH, ALRP, AR and iNOS expression was decreased in EH, AR. 5. PGE2 and COX-2 expression was decreased in EH, AR. 6. $IL-1{\beta}$ production was significantly decreased in EH, AR and IL-6 production was significantly decreased in AR. $TNF-{\alpha}$ production was significantly decreased in ALRP, AR. 7. MMP-9 and TIMP-1 production were significantly decreased in EH. 8. HO-1 expression was significantly increased in EH. 9. With simultaneous usage of SnPP which is expression inhibitor of HO-1, NO, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production were partially increased in EH, ALRP, AR. Conclusions According to this study, Components of Mahwangbujaseshin-tang have anti-oxidants and anti-inflammation effects in LPS-Stimulated RAW264.7 Macrophages.

• Archives of Pharmacal Research
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• 제27권3호
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• pp.291-294
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• 2004

Activity-guided fractionation of the n-hexane and ${CHCl_3}-soluble$ fractions of Sindora sumatrana using a bioassay based on the inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production by inducible nitric oxide synthase (iNOS) in murine macrophage RAW 264.7 cells led to the isolation of the known compound, $(+)-7{\beta}-acetoxy-15,16-epoxy-3$, 13(16), 14-clero-datriene-18-oic acid (2) as an active constituent. In addition, a new trans-clerodane diterpenoid, (+)-2-oxokolavenic acid (1), together with six known compounds, (+)-3, 13-clerodadiene-16,15-olide-18-oic acid (3), $(+)-7{\beta}-acetoxy-3$,13-clerodadiene-16,15-olide-18-oic acid (4), $(+)-7{\beta}-acetoxy-16-hydroxy-3$,13-clerodadiene-16, 15-olide-18-oic acid (5), ${\beta}-caryophyllene$ oxide (6), $clovane-2{\beta},9{\beta}-diol (7),{\;}and{\;}caryolane-1,9{\beta}-diol$ (8) were isolated and found to be inactive. The structure of compound 1 was determined using physical and spectroscopic methods such as 1D and 2D-NMR experiments. The known compounds 2-8 were identified by the spectroscopic data and by comparison with the published values. Of eight isolates (1-8), only compound 2 exhibited an iNOS inhibitory activity with $IC_{50}$/ value of $51.6{\;}\mu\textrm{m}M$.

• Biomolecules & Therapeutics
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• 제22권5호
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• pp.390-399
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• 2014

Chalcones (1,3-diaryl-2-propen-1-ones), a flavonoid subfamily, are widely known for their anti-inflammatory properties. Propenone moiety in chalcones is known to play an important role in generating biological responses by chalcones. In the present study, we synthesized chalcone derivatives structurally modified in propenone moiety and examined inhibitory effect on nitric oxide (NO) production and its potential mechanisms. Among the chalcone derivatives used for this study, TI-I-174 (3-(2-Hydroxyphenyl)-1-(thiophen-3-yl)prop-2-en-1-one) most potently inhibited lipopolysaccharide (LPS)-stimulated nitrite production in RAW 264.7 macrophages. TI-I-174 treatment also markedly inhibited inducible nitric oxide synthase (iNOS) expression. However, TI-I-174 did not significantly affect production of IL-6, cyclooxygenase-2 (COX-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that TI-I-174 inhibits production of inflammatory mediators in a selective manner. Treatment of macrophages with TI-I-174 significantly inhibited transcriptional activity of activator protein-1 (AP-1) as determined by luciferase reporter gene assay, whereas nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not affected by TI-I-1744. In addition, TI-I-174 significantly inhibited activation of c-Jun-N-Terminal kinase (JNK) without affecting ERK1/2 and p38MAPK, indicating that down-regulation of iNOS gene expression by TI-I-174 is mainly attributed by blockade of JNK/AP-1 activation. We also demonstrated that TI-I-174 treatment led to an increase in heme oxygenase-1 (HO-1) expression both at mRNA and protein level. Transfection of siRNA targeting HO-1 reversed TI-I-174-mediated inhibition of nitrite production. Taken together, these results indicate that TI-I-174 suppresses NO production in LPS-stimulated RAW 264.7 macrophages via induction of HO-1 and blockade of AP-1 activation.

• IMMUNE NETWORK
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• 제10권6호
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• pp.212-218
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• 2010

Backgroud: The stem bark of Kalopanax pictus (KP) has been used in traditional medicine to treat rheumatoidal arthritis, neurotic pain and diabetes mellitus in China and Korea. In this study, the mechanism responsible for anti-inflammatory effects of KP was investigated. Methods: We examined the effects of KP on NO production, nitric oxide synthase (iNOS) and HO-1 expression, NF-${\kappa}B$, Nrf2 and MAPK activation in mouse peritoneal macrophages. Results: The aqueous extract of KP inhibited LPS-induced NO secretion as well as inducible iNOS expression, without affecting cell viability. KP suppressed LPS-induced NF-${\kappa}B$ activation, phosphorylation and degradation of $I{\kappa}B-{\alpha}$, phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun N-terminal kinase (JNK). Furthermore, KP induced HO-1 expression and Nrf2 nuclear translocation. Conclusion: These results suggest that KP has the inhibitory effects on LPS-induced NO production in macrophages through NF-${\kappa}B$ suppression and HO-1 induction.

• Proceedings of the Korean Society of Applied Pharmacology
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• 한국응용약물학회 2004년도 Annual Meeting of KSAP : New Drug Development from Natural Products
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• pp.3-10
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• 2004

Oxidative stress is a constant threat to all living organisms and an immense repertoire of cellular defense systems is being employed by most pro- and eukaryotic systems to eliminate or to attenuate oxidative stress. Ischemia and reperfusion is characterized by both a significant oxidative stress and characteristic changes in the antioxidant defense. Heme oxigenase-l (HO-l) is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against ischemic injury in mammalian cells. Higenamine, an active ingredient of Aconite tuber, has been shown to have antioxidant activity along with inhibitory action of inducible nitric oxide synthase (iNOS) expression in various cells. In the present study, we investigated whether higenamine and related analogs protect cells from oxidative cellular injuries by modulating antioxidant enzymes, such as HO-l, MnSOD etc. R-form of YS-51 was the most potent inducer of HO-l in bovine endothelial cells, which inhibited apoptotic cell death by H$_2$O$_2$. HO-1 induction by YS 51 was mediated by PI3 kinase activation in which PKA- as well as PKG pathway is considered as important regulators. YS-51 also induced Mn-SOD mRNA expression by activating c-jun N-terminal kinase in endothelial cells and Hela cells. In ROS 17/2.1 cells, higenamine and enetiomers of related compounds inhibited iNOS expression by cytokine mixtures. Taken together, higenamine and related compounds can be developed as possible protective agents from oxidative cell injury or death.

• Journal of Korean Medicine Rehabilitation
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• 제23권2호
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• pp.1-15
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• 2013

Objectives : In present study, we investigated the effects of ginsenoside Rg3 on early-stage inflammatory response in spinal cord compression of rodents. Methods : Spinal cord injury(SCI) was induced by a vascular clip method(30 g, 5 min) on the spinal cord of mice. Rg3 was treated orally at 1 hour prior to the SCI induction. Messenger ribonucleic acid(mRNA) expression of tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), interleukin-1${\beta}$(IL-1${\beta}$), interleukin-6(IL-6) and cyclooxygenase-2(COX-2) was measured by the real-time polymerase chain reaction(RT-PCR). Microglia in the spinal cord tissue, neurophils and COX-2 in the peri-lesion and inducible nitric oxide synthase(iNOS) expression in the ventral horn of SCI induced rats were measured by immunohistochemical stain. Results : 1. Rg3 significantly reduced the mRNA expression of TNF-${\alpha}$, IL-1${\beta}$, and COX-2 in the spinal cord tissue compared with SCI group(p<0.05, p<0.01). 2. Rg3 significantly reduced the total number of activated microglia and proportion of phagocytic form in the total activated microglia compared with SCI group(p<0.05, p<0.01). 3. Rg3 significantly reduced myeloperoxidase(MPO) positive neurophil in the peri-lesion compared with SCI group(p<0.05). 4. Rg3 reduced the COX-2 expression in the tissue and motor neurons compared with SCI group. 5. Rg3 significantly reduced iNOS positive motor neurons in the ventral horn compared with SCI group(p<0.01). Conclusions : In conclusion, we demonstrated at first that treatment of ginsenoside Rg3 could reduce significantly the levels of inflammatory mediators in a spinal cord compression model of rodents. Therefore, these results suggested that ginsenoside Rg3 may be a useful antimiflamatory therapeutic candidate for SCI.

• The Korea Journal of Herbology
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• 제31권5호
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• pp.1-6
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• 2016

Objectives : Nardostachys jatamansi (NJ) is a medicinal herb that has been reported in various traditional systems of medicine for its use in antispasmodic, a digestive stimulant, skin diseases. Previous studies have already reported that NJ effectively protects against inflammation. However, the active compound in NJ is unknown. Therefore, in the present study, we analyzed effects of a compound, 8α-hydroxy pinoresinol (HP), isolated from NJ against lipopolysaccharide (LPS) induced inflammation in RAW 264.7 cells.Methods : To examine the anti-inflammatory effect of HP against LPS, intraperitoneally pre-treat the HP (100, 200, 500 and 1,000 nM) 1 h prior to LPS challenges. LPS was stimulated with 500 ng/ml in RAW 264.7 cells. To identify the anti-inflammatory effect of HP, we measured inflammatory mediators such as inducible nitric oxide synthase (iNOS) and its derivative nitric oxide (NO), cyclooxygenase-2 (COX-2), prostaglandin E2 (PGE2). Also we evaluated molecular mechanisms including mitogen-activated protein kinases (MAPKs) and nuclear factor-kappaB (NF-κB) activation by western blot.Results : The HP inhibited production of inflammatory mediators, such as iNOS and its derivative NO, COX-2 and PGE2 in LPS- induced inflammationin RAW 264.7 cells. Additionally, HP also inhibited activation of p38 pathway signaling but not extracellularsignal-regulatedkinase (ERK), c-jun NH2-terminal kinase (JNK), and NF-κB.Conclusion : Our results suggest that HP has anti-inflammatory functions through the dephosphorylation of p38 and HP can provide beneficial strategy for prevention and therapy of inflammation.

• Preventive Nutrition and Food Science
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• 제15권3호
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• pp.206-212
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• 2010

Codium fragile (CF) is an edible green alga consumed as a traditional food source in Korea. In this study, the ethanol extract of CF was evaluated to determine if it has anti-inflammatory activity. Lipopolysaccharide (LPS), a toxin from bacteria, is a potent inducer of inflammatory cytokines, such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Therefore, we studied whether CF extracts have an anti-inflammatory effect in LPS-induced murine macrophage cell lines (RAW 264.7). In the present study, IL-6 production was measured using an enzyme-linked immunosorbent assay (ELISA), prostaglandin $E_2$($PGE_2$) production was measured using the EIA kit, and cyclooxygenase (COX)-2 and mitogen-activated protein kinase (MAPK) activation were determined by Western blot analysis. IL-6 mRNA, COX-2 mRNA and iNOS mRNA expression were measured using reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that CF extracts inhibit LPS-induced IL-6, NO and PGE2 production in a dose-dependent manner, as well as expression of iNOS and COX-2. CF extracts significantly inhibited LPS-induced c-Jun N-terminal kinase (JNK) 1/2 phosphorylation. Taken together, these findings may help elucidate the mechanism by which CF modulates RAW 264.7 cell activation under inflammatory conditions.

• The Korean Journal of Physiology and Pharmacology
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• 제11권1호
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• pp.9-13
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• 2007

Antioxidant properties have been proposed as a mechanism for the putative anti-inflammatory effects of phenolic compounds. To reveal the relationship between antioxidant activity and anti-inflammatory effects of various antioxidants, we measured 1, 1-diphenyl-2-picryhydrazyl(DPPH)-reducing activity and examined the inhibitory effects on LPS-induced inflammation-related gene expression in the BV2 microglial cell line. Lipopolysaccharide(LPS)(0.2 ${\mu}g/ml$) was used with or without antioxidants to treat cells, and the regulation of iNOS and cytokine gene expression was monitored using an RNase protection assay(RPA). Although, all tested antioxidants had similar DPPH-reducing activity and inhibited nitrite production, but the curcuminoid antioxidants(ferulic acid, caffeic acid, and curcumin) inhibited LPS-induced gene expression(iNOS, $TNF-\alpha,\;IL-1{\beta}$, IL-6, and IL-1 Ra) in a concentration-dependent manner. Other tested antioxidants did not exhibit the same effects; N-acetylcysteine(NAC) only began to suppress $IL-1{\beta}$ gene expression just below the concentration at which cytotoxicity occurred. Moreover, the antioxidant potency of curcuminoids appeared to have no correlation with anti-inflammatory potency. Only curcumin could inhibit LPS-induced microglial activation at a micromolar level. These data suggest that curcumin may be a safe antioxidant possessing anti-inflammatory activity.

• The Korean Journal of Physiology and Pharmacology
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• 제24권5호
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• pp.395-402
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• 2020

This study has investigated the effect of a potent bioflavonoid, troxerutin, on diabetes-induced changes in pro-inflammatory mediators and expression of microRNA-146a and nuclear factor-kappa-B (NF-κB) signaling pathway in aortic tissue of type-I diabetic rats. Male Wistar rats were randomly divided into four groups (n = 6/each): healthy, healthy-troxerutin, diabetic, and diabetic-troxerutin. Diabetes was induced by streptozotocin injection (60 mg/kg; intraperitoneally) and lasted 10 weeks. Troxerutin (150 mg/kg/day) was administered orally for last month of experiment. Inflammatory cytokines IL-1β, IL-6, and TNF-α, as well as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule (VCAM), cyclooxygenase-II (COX-II), and inducible-nitric oxide synthase (iNOS) were measured on aortic samples by enzyme-linked immunosorbent assay. Gene expressions for transcription factor NF-κB, interleukin-1 receptor-associated kinase-1 (IRAK-1), TNF receptor-associated factor-6 (TRAF-6), and microRNA-146a were determined using real-time polymerase chain reaction. Ten-week diabetes significantly increased mRNA levels of IRAK-1, TRAF-6, NF-κB, and protein levels of cytokines IL-1β, IL-6, TNF-α, adhesion molecules ICAM-1, VCAM, and iNOS, COX-II, and decreased expression of microRNA-146a as compared with healthy rats (p < 0.05 to p < 0.01). However, one month treatment of diabetic rats with troxerutin restored glucose and insulin levels, significantly decreased expression of inflammatory genes and pro-inflammatory mediators and increased microRNA level in comparison to diabetic group (p < 0.05 to p < 0.01). In healthy rats, troxerutin had significant reducing effect only on NF-κB, TNF-α and COX-II levels (p < 0.05). Beside slight improvement of hyperglycemia, troxerutin prevented the activation of NF-κB-dependent inflammatory signaling in the aorta of diabetic rats, and this response may be regulated by microRNA-146a.