• Korean Journal of Organic Agriculture
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• v.28 no.4
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• pp.591-604
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• 2020

The objectives of this study were to find out the best condition of extracting methods of limonene from citron and to determine effects of limonene on immune modulation activity by measuring cytokine secretion using RAW 264.7 mouse macrophage cells. When distilled water was used as a solvent instead of organic solvents to extract limonene from citron, addition of refluxing process to simultaneous steam distillation extraction method was found to be much effective in extracting limonene. However, it required longer extraction time than using other organic solvents. Limonene extracts showed increased IL-β and IL-6 but decreased the TNF-α gene expression in limonene concentration dependant manner. However oral administration of limonene extracts to mice did not influence significantly compared to control in in vivo experiment. It might be due to that the mice were kept in well controlled and complete environment. Limonene, a natural material from citron has been approved to have a immune-modulation activity in the present study and have a potential as a feed additive that is environmentally friendly and no harmful. Further study with protected limonene, for example, for the protection of limonene from oxidation or bypass the ruminal degradation in order consequently to increase immune-modulation activity might be useful as a further research.

• Journal of Ginseng Research
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• v.43 no.3
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• pp.377-384
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• 2019

Background: Inflammation is widespread in the clinical pathology and closely associated to the progress of many diseases. Triterpenoid saponins as a key group of active ingredients in Panax notoginseng (Burk.) F.H. Chen were demonstrated to show antiinflammatory effects. However, the chemical structures of saponins in the leaves and stems of Panax notoginseng (PNLS) are still not fully clear. Herein, the isolation, purification and further evaluation of the antiinflammatory activity of dammarane-type triterpenoid saponins from PNLS were conducted. Methods: Silica gel and reversed-phase C8 column chromatography were used. Furthermore, preparative HPLC was used as a final purification technique to obtain minor saponins with high purities. MS, NMR experiments, and chemical methods were used in the structural identifications. The antiinflammatory activities of the isolated saponins were assessed by measuring the nitric oxide production in RAW 264.7 cells stimulated by lipopolysaccharides. Real-time reverse transcription polymerase chain reaction was used to measure the gene expressions of inflammation-related gene. Results: Eight new minor dammarane-type triterpene oligoglycosides, namely notoginsenosides LK1-LK8 (1-8) were obtained from PNLS, along with seven known ones. Among the isolated saponins, gypenoside IX significantly suppressed the nitric oxide production and inflammatory cytokines including tumor necrosis $factor-{\alpha}$, interleukin 10, interferon-inducible protein 10 and $interleukin-1{\beta}$. Conclusion: The eight saponins may enrich and expand the chemical library of saponins in Panax genus. Moreover, it is reported for the first time that gypenoside IX showed moderate antiinflammatory activity.

• Journal of the Korean Applied Science and Technology
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• v.38 no.5
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• pp.1302-1313
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• 2021

In this study, Gami-Sumiwon (GS; Cynanchi wilfordii R., Angelica gigantis R., Lycium chinense M., Betula platyphylla S., Cryptotympana atrata F. and Carthamus tinctorius L.) and/or Sinomenium acutum R. (SC) was extracted with 70% ethanol or water. And We investigated the antioxidant activity effect of GS±SC. The following experimental techniques were used to evaluate the antioxidant efficacy of GS±SC. HPLC chromatogram, heavy metal content, ABTS/DPPH radical scavenging analysis, SOD-like activity assay, FACS, and NO assay. As a result of the experiment, the sinomenine content was found to be higher in DW extracts, and decursin was found to be higher in 80% ethanol extracts. And, the amount of heavy metals in all extracts was below the standard value. ABTS, and DPPH radical scavenging activity was identified that GS±SC(EtOH) was found to have a higher scavenging activity than GS±SC(DW). But, SOD showed the opposite result. No cytotoxicity of GS was observed on Raw 264.7 cells at concentration of 1~100 ㎍/㎖. The ROS production was significantly decreased that GS±SC(DW) was found to have a higher scavenging activity than GS±SC(EtOH). However, NO production showed the opposite result. Looking at the results of SOD and ROS analysis, SC does not seem to have a function of prevention. SC is thought to have an effect on the removal of free radicals generated after oxidative stress. This result objectively confirmed the antioxidant effect of GS±SCs. We will continue to conduct in-depth research. Therefore, it is believed that the possibility of using GS±SCs as a functional material can be established. The more diverse the objectives, the higher the value of GS utilization is thought to be.

• Korean Journal of Food Science and Technology
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• v.54 no.2
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• pp.147-154
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• 2022

The Mongolian lingonberry and blueberry are two essential food sources found in Mongolia. This study investigated the antioxidant and anti-inflammatory effects of methanol extracts from Mongolian lingonberry (LBE) and blueberry (BBE). Compared to the LBE, the BBE showed higher total phenolic, flavonoid, and anthocyanin contents, as well as antioxidant capacities. The LBE and BBE inhibited the mRNA expression of pro-inflammatory genes, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and cyclooxygenase (COX-2) in lipopolysaccharide-stimulated RAW 264.7 macrophage cells. In addition, the LBE and BBE inhibited NADPH oxidase-2 (Nox2) mRNA expression, indicating that they have cellular antioxidant capacities. Anthocyanin derivatives of the LBE and BBE were analyzed using LC-QTOF/MS. Six anthocyanins were identified in the BBE, while one was detected in the LBE. Our findings demonstrate that the anthocyanin-rich LBE and BBE could be used as functional food sources in Mongolia.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.36 no.1
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• pp.21-39
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• 2023

Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Goihwa-san water extract(GHS) in vitro & in vivo. Methods : In vitro, we evaluated the anti-inflammatory effect of GHS by comparing the Raw 264.7 cells with 10, 30, 100, 300㎍/㎖ of GHS for 1 hour before Lipopolysaccharide(LPS) to the single LPS treated group. We examined the relative cell viability by MTT assay and the relative level of LPS, Loxoribine(LOX), Peptidoglycan(PGN), Flagellin(FLA)-induced NO production by using Griess reagent and measured relative iNOS protein level and COX-2 protein level by using western blot and Image analyzing system. We measured the production of TNF-α, IL-1β, and IL-6 by each ELISA kits and then measured the relative levels of IκBα, p-IκBα in whole-cell lysate fraction and NF-κB in nuclear fraction by using western blot and Image analyzing system. In vivo, we induced the paw edema by subcutaneous injection of 100㎕/rat CA and measured the swelling volume of paw by using a plethysmometer and then measured the relative iNOS protein level by using western blot. Results : As a result, in vitro, LPS, PGN-induced NO production was significantly inhibited by pretreatment with GHS. GHS reduced LPS, PGN-induced iNOS expression, PGN-induced COX-2 expression and LPS-induced production of cytokine(TNF-α, IL-1β, IL-6). Expression of IκBα was increased by pretreatment with GHS 100㎍/㎖. And the expression of p-IκBα and NF-κB were decreased by pretreatment with GHS 100㎍/㎖. In vivo, CA-induced inflammation rat model was used for the evaluation of the anti-inflammatory effect of GHS. 0.3 or 1.0g/kg of GHS significantly reduced the increases of paw swelling and iNOS expression in paw tissues. Conclusions : These results show that GHS can decrease inflammatory response via inhibition of the NF-κB pathway in vitro. And in vivo, the anti-inflammatory effect suggest the clinical basis of GHS for the treatment of inflammatory diseases.

• The Korea Journal of Herbology
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• v.32 no.3
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• pp.45-53
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• 2017

Objective : Nelumbo nucifera, its rhizome and semen have been used as a traditional medicine which was studied on antioxidant, hepatoprotective effect, anti-obesity and the others. However, Nelumbinis Flos have not studied. We investigated protective effects on oxidative DNA damage and anti-inflammatory effects of Nelumbinis Flos. Methods : The antioxidant activity was conducted by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay and reducing power assay. Total phenolic content was analyzed. Also, phenolic compounds were detected by HPLC/UV. The inhibitory effect on oxidative DNA damage was determined using ${\Phi}X-174$ RF I plasmid DNA cleavage assay. The anti-inflammatory effect of Nelumbinis Flos was measured by the amount of nitric-oxide (NO) produced and protein levels of iNOS, and COX-2 in LPS induced RAW 264.7 cells. Results : The results of DPPH and ABTS radical scavenging activity at $200{\mu}g/m{\ell}$ of extraction were $97.02{\pm}0.88%$ and $96.42{\pm}0.25%$. Reducing power (fold of L-ascorbic acid as control) was $100.14{\pm}0.31$ at $200{\mu}g/m{\ell}$. Total phenol content was $8.70{\pm}0.02mg/g$. Chlorogenic acid, catechin and epicatechin were found by HPLC. Nelumbinis Flos has inhibitory effect in dose-manner against oxidative DNA damage. In addition, it showed the anti-inflammatory effect by suppression of NO production as well as protein levels of iNOS, and COX-2. Conclusion : This study suggested that Nelumbinis Flos showed potential antioxidant and suppression activities of various factors were related in NO produced. Therefore, Nelumbinis Flos as natural plant resources that may help reduce inflammation and alleviate DNA damage.

• Korean Journal of Clinical Laboratory Science
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• v.54 no.3
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• pp.209-216
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• 2022

The effect of Saccharomycopsis fibuligera fermentation on the antioxidant and anti-inflammatory activity of Kerria japonica (K. japonica) extracts was studied. First, the antioxidant activity of the fermented extract was measured using the 2,2-diphenyl1-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzthiazoline 6-sulfonic acid (ABTS) methods. Also, the quantification of polyphenols and flavonoids, which are representative components with antioxidant activity, was performed. The results of the DPPH and ABTS assays showed an increase in the antioxidant activity by 14.39% and 21.74%, respectively, due to fermentation. The polyphenol concentration increased by 10.5%, and the flavonoid concentration increased by 100.0%. In the cell experiment, a cytotoxicity test and a nitric oxide (NO) production inhibitory test were performed using RAW 264.7 cells. Both the control group and the fermentation group showed no cytotoxicity. In the NO production inhibition experiment, the fermentation group showed a 6.85% higher inhibition of NO production compared to the control group. When the inhibitory effects of the extracts on inflammatory cytokine production were assessed, the fermentation group showed 12.4% and 23.5% higher inhibition of interleukin (IL)-1β and IL-6 production, respectively, compared to the control group. In conclusion, due to its potential for inhibiting NO and inflammatory cytokine production, fermented K. japonica extracts could be considered a source of anti-inflammatory compounds.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.39-50
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• 2009

Objectives : Smilacis glabrae rhizoma (SG) has been traditionally used as a herbal medication of musculoskeletal disorders like arthritis, pain, convulsions, and syphilis in traditional Korean medicine. This study was investigated anti-oxidative and anti-inflammatory effect of fractionated extracts of Smilacis Glabrae Rhizoma in Human Umbilical Vein Endothelial Cell (HUVEC). Methods : SG extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of SG onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymethoxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of ICAM-1 and VCAM-1 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially ethyl acetate (EA) extract, significantly inhibited free radical generation, the TNF-$\alpha$-induced intracellular oxidation. Furthermore, the EA extract protected TNF-$\alpha$-induced adhesion to THP-1, expression of adhesion molecules accompanied by an attenuation of IL-6 and IL-8 formation in HUVEC. Conclusions : These results indicate that EA extract of SG have potential as an agent of atherosclerosis and other chronic inflammatory diseases including diabetes, hypertension, and arthritis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.6
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• pp.819-827
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• 2016

This study was carried out to investigate anti-inflammatory, anti-diabetic, alcohol metabolizing, and hepatoprotective effects of hot water (MOW) and 80% ethanol (MOE) extracts from moringa (Moringa oleifera Lam.) leaf. The total phenol content of MOW and MOE were 45.49 and 63.06 mg tannic acid equivalents/g, respectively. 1,1-Diphenyl-2-picrylhydrazyl radical scavenging activities of MOW and MOE were remarkably elevated in a dose-dependent manner, and about 60.8% and 71.3% at 1 mg/mL, respectively (P<0.01). Superoxide dismutase-like activities of MOW and MOE were 2.8% and 7.4% at 5 mg/mL, respectively (P<0.05). ${\alpha}-Glucosidase$ inhibitory activity also increased in a dose-dependent manner in both extracts, and MOE was higher about two times than MOW at 5 mg/mL (P<0.001). The effects of MOW and MOE on alcohol metabolizing activity were determined by measuring generation of reduced nicotinamide adenine dinucleotide (NADH) by alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH). ADH and ALDH activities significantly increased upon addition of MOW and MOE (P<0.05). Anti-inflammatory activity was examined in lipopolysaccharide-stimulated RAW 264.7 cells. Nitric oxide production was reduced to 32.1% and 81.2% by addition of MOW and MOE at 1 mg/mL, respectively (P<0.05). MOW and MOE showed significant protective effects against tacrine-induced cytotoxicity in Hep3B cells at $100{\mu}g/mL$. These results suggest that moringa leaf extracts have great potential as natural health products.

• Journal of Veterinary Clinics
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• v.28 no.2
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• pp.190-195
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• 2011

Nuclear factor ${\kappa}B$ (NF-${\kappa}B$) is a nuclear transcription factor that modulates the expression of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. trans-10, cis-12 (t10c12)-conjugated linoleic acid (CLA) participates in the inhibition of TNF-${\alpha}$ production upon lipopolysaccharide (LPS)-stimulation. However, in our previous study, t10c12-CLA enhanced the production of TNF-${\alpha}$ by LPS-unstimulated porcine peripheral blood mononuclear cells (PBMCs) and RAW 264.7 macrophages in vitro. To resolve this apparent contradiction, we hypothesized that the effect of t10c12-CLA on TNF-${\alpha}$ production depends on NF-${\kappa}B$ activation induced by LPS stimulation. To test this hypothesis, we assessed the in vitro effect of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ p65 activity in LPS-stimulated and LPS-unstimulated porcine PBMCs. t10c12-CLA treatment resulted in increased TNF-${\alpha}$ production by LPS-unstimulated PBMCs but decreased TNF-${\alpha}$ production by LPS-stimulated PBMCs. t10c12-CLA increased the degradation of inhibitory ${\kappa}B$ ($I{\kappa}B$)-${\alpha}$ protein and activated NF-${\kappa}B$ p65 in LPS-unstimulated PBMCs, but had the opposite effect in LPS-stimulated PBMCs. Notably, t10c12-CLA enhanced NF-${\kappa}B$ p65 binding activity in LPS-unstimulated PBMCs exposed to caffeic acid phenethyl ester (CAPE), a NF-${\kappa}B$ inhibitor. Conversely, it inhibited NF-${\kappa}B$ p65 binding activity in LPS-stimulated PBMCs exposed to CAPE. These results suggest that t10c12-CLA may have different actions under different physiological conditions, and that its effect may be associated with a change in NF-${\kappa}B$ p65 activity.