• Nutrition Research and Practice
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• v.11 no.1
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• pp.3-10
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• 2017

BACKGROUND/OBJECTIVES: Sargassum horneri is an edible brown alga that grows in the subtidal zone as an annual species along the coasts of South Korea, China, and Japan. Recently, an extreme amount of S. horneri moved into the coasts of Jeju Island from the east coast of China, which made huge economic and environmental loss to the Jeju Island. Thus, utilization of this biomass becomes a big issue with the local authorities. Therefore, the present study was performed to evaluate the anti-inflammatory potential of crude polysaccharides (CPs) extracted from S. horneri China strain in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. MATERIALS/METHODS: CPs were precipitated from S. horneri digests prepared by enzyme assistant extraction using four food-grade enzymes (AMG, Celluclast, Viscozyme, and Alcalase). The production levels of nitric oxide (NO) and pro-inflammatory cytokines, including tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-$1{\beta}$ were measured by Griess assay and enzyme-linked immunosorbent assay, respectively. The levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), nuclear factor (NF)-${\kappa}B$, and mitogen-activated protein kinases (MAPKs) were measured by using western blot. The IR spectrums of the CPs were recorded using a fourier transform infrared spectroscopy (FT-IR) spectrometer. RESULTS: The polysaccharides from the Celluclast enzyme digest (CCP) showed the highest inhibition of NO production in LPS-stimulated RAW 264.7 cells ($IC_{50}$ value: $95.7{\mu}g/mL$). Also, CCP dose-dependently down-regulated the protein expression levels of iNOS and COX-2 as well as the production of inflammatory cytokines, including TNF-${\alpha}$ and IL-$1{\beta}$, compared to the only LPS-treated cells. In addition, CCP inhibited the activation of NF-${\kappa}B$ p50 and p65 and the phosphorylation of MAPKs, including p38 and extracellular signal-regulated kinase, in LPS-stimulated RAW 264.7 cells. Furthermore, FT-IR analysis showed that the FT-IR spectrum of CCP is similar to that of commercial fucoidan. CONCLUSIONS: Our results suggest that CCP has anti-inflammatory activities and is a potential candidate for the formulation of a functional food ingredient or/and drug to treat inflammatory diseases.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.38-45
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• 2018

Acne is an inflammatory skin disease that occurs in puberty and young people. Propionibacterium acnes (P. acnes) is known to be a major cause of inflammation in acne. P. acnes proliferates within hair follicles blocked by overproduced sebum in the skin, and thereby activates monocytic cells to promote the secretion of pro-inflammatory cytokines. In this study, we investigated the possibility of Cnestis palala (Lour.) Merr. extract to diminish P. acnes-mediated inflammatory responses. We found that C. palala extract significantly attenuated P. acnes-induced pro-inflammatory cytokine expressions, such as $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, iNOS, and COX-2 in mouse macrophage RAW264.7 cells. Moreover, we observed that C. palala extract inhibited $NF-{\kappa}B$ transcriptional activation, which is the major transcription factor of inflammatory cytokine expression. Therefore, it is expected that C. palala extract has a potential as a therapeutic agent or supplement for the treatment P. acnes-induced inflammatory responses.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.1
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• pp.7-13
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• 2015

In this study, anti-inflammatory activity of hot water extract of Aronia fruits (AF-H) was examined. Pre-treatment with AF-H significantly inhibited production of nitric oxide (NO) and prostaglandin E-2 in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The inhibitory effect of AF-H on LPS-induced inflammation was also confirmed by down-regulation of inducible NO synthase as well as cyclooxygenase-2 protein expression. Furthermore, treatment with AF-H significantly inhibited secretion of inflammatory cytokines such as tumor-necrosis $factor-{\alpha}$ and interleukin-6. Signal transduction pathway studies further indicated that AF-H inhibited LPS-induced activation of nuclear $factor-{\kappa}B$, but not mitogen-activated protein kinase. Treatment with AF-H also partially protected against LPS-induced lethal shock in C57BL/6 mice, although its effect was not statistically significant. These results suggest that AF-H is a more promising nutraceutical or medicinal agent for inhibition of LPS-induced inflammation or inflammation-related diseases.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.11
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• pp.1776-1788
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• 2019

Objective: The demands for measures to improve disease resistance and productivity of livestock are increasing, as most countries prohibit the addition of antibiotics to feed. This study therefore aimed to uncover functional feed additives to help enhance livestock immunity and disease resistance, using Acanthopanax sessiliflorus fruit extract (ASF). Methods: ASF was extracted with 70% EtOH, and total polyphenolic and catechin contents were measured by the Folin-Ciocalteu and vanillin assay, respectively. The 3D4/31 porcine macrophage cells ($M{\Phi}$) were activated by phorbol 12-myristate 13-acetate (PMA), and cell survival and growth rate were measured with or without ASF treatment. Flow-cytometric analysis determined the lysosomal activity, reactive oxygen species levels (ROS), and cell cycle distribution. Nuclear factor kappa B ($NF-{\kappa}B$) and superoxide dismutase (SOD) protein expression levels were quantified by western blotting and densitometry analysis. Quantitative polymerase chain reaction was applied to measure the lipid metabolism-related genes expression level. Lastly, the antibacterial activity of 3D4/31 $M{\Phi}$ cells was evaluated by the colony forming unit assay. Results: ASF upregulated the cell viability and growth rate of 3D4/31 $M{\Phi}$, with or without PMA activation. Moreover, lysosomal activity and intracellular ROS levels were increased after ASF exposure. In addition, the antioxidant enzyme SOD2 expression levels were proportionately increased with ROS levels. Both ASF and PMA treatment resulted in upregulation of $NF-{\kappa}B$ protein, tumor necrosis factor $(TNF){\alpha}$ mRNA expression levels, lipid synthesis, and fatty acid oxidation metabolism. Interestingly, co-treatment of ASF with PMA resulted in recovery of $NF-{\kappa}B$, $TNF{\alpha}$, and lipid metabolism levels. Finally, ASF pretreatment enhanced the in vitro bactericidal activity of 3D4/31 $M{\Phi}$ against Escherichia coli. Conclusion: This study provides a novel insight into the regulation of $NF-{\kappa}B$ activity and lipid metabolism in $M{\Phi}$, and we anticipate that ASF has the potential to be effective as a feed additive to enhance livestock immunity.

• Korean Journal of Plant Resources
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• v.32 no.5
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• pp.433-441
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• 2019

This study describes a preliminary evaluation of the anti-inflammatory activity and anti-atopic activity of Phormium tenax leaf extracts. P. tenax leaf was extracted using 70% ethanol and then fractionated sequentially with n-hexane, methylene chloride, ethyl acetate, n-butanol. In order to effectively screen for anti-inflammatory agents, we first investigated the inhibitory effects of P. tenax leaf crude extracts and solvent fractions on production of pro-inflammatory factors[nitric oxide(NO), prostaglandin $E_2(PGE_2)$, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2)] and pro-inflammatory cytokines [tumor necrosis $factor-{\alpha}(TNF-{\alpha})$, interleukin-6(IL-6) and $interleukin-1{\beta}(IL-1{\beta})$] in lipopolysaccharide(LPS)-stimulated RAW 264.7 cells. In addition, we also evaluated of their inhibitory effect on the atopic dermatitis-like inflammatory markers such as macrophage-derived chemokine(MDC) and thymus and activation-regulated chemokine(TARC) in HaCaT cells. Among the five solvent fractions of P. tenax, methylene chloride and ethyl acetate fractions inhibited production of pro-inflammatory factors and pro-inflammatory cytokines in a dose dependent manner, respectively. These fractions were also showed inhibitory activity for MDC and TARC expression levels in $IFN-{\gamma}-stimulated$ HaCaT cells, respectively. These results suggest that P. tenax have significantly effects of anti-inflammatory activity and anti-atopic activity that might be beneficial for the topical treatment of inflammatory skin disorders.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.8
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• pp.477-484
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• 2019

Several types of scavenger receptors, including the Collectin-Placenta 1 (CL-P1) receptorthat is present in mammals, are molecules that are expressed on the surfaces of endothelial cells, macrophages and smooth muscle cells. These molecules are cell surface glycoproteins that can be conjugated to oxidized low density lipoprotein (oxLDL). Among these molecules, the effect of quercetin on CL-P1 activation has been confirmed. Quercetin is known as an antioxidant that stops oxidation because it acts to remove free radicals that are responsible for the oxidation reaction. In this study, fragments from the transcription start site of the mouse CL-P1 gene promoter to the -500th base were cloned using DNA polymerase. These fragments were then introduced into macrophage like RAW264.7 cells and fibroblast-like NIH3T3 cells to study the effect of quercetin on the CL-P1 gene expression. As a result, we found that bases ranging from -250 to -350 in the anterior part where gene expression starts are important for producing CL-P1 protein. Among them, the DNA mutation experiments we performed confirmed that the E2F binding sites are critical for producing the CL-P1 protein? In addition, when quercetin was added to the RAW264.7 culture medium, which was a culture of adherent cells, observedthe phenomenon of the cells falling off from the surface of the culture container.

• Korean Journal of Microbiology
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• v.55 no.3
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• pp.213-219
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• 2019

Acne is a chronic inflammatory disease outbreak in the sebaceous glands within the hair follicle. The proliferation of Propionibacterium acnes (P. acnes) causes monocytes to stimulate secretion of inflammatory cytokines. A number of studies proposed the inhibitory effects of P. acnes-mediated inflammation by several natural extracts. However, studies on the effect of Eurya persicifolia Gagnep. (E. persicifolia) extracts on the inflammatory responses by P. acnes have not been explored yet. In this study, we investigated the anti-inflammatory effect of E. persicifolia extract in the inflammatory reactions induced by P. acnes. We found that E. persicifolia extract successfully diminished the expression levels of inflammatory mediators such as IL-$1{\beta}$, IL-6, TNF-${\alpha}$, and iNOS in P. acnes-activated mouse macrophage RAW 264.7 cells. We found that the immunosuppressive effect of E. persicifolia extract in the P. acnes-activated inflammatory signaling is mediated by the regulation of NF-${\kappa}B$ transcriptional activation, which is a key regulator of inflammatory cytokine expression. Our results suggest that E. persicifolia extract held potentials for the treatment of P. acnes by suppressing NF-${\kappa}B$ signaling pathways.

• Journal of Life Science
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• v.31 no.8
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• pp.745-754
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• 2021

Deterioration of the immune function weakens the body's resistance to various infections, leading to a series of diseases. Immunomodulatory biomaterials have been used to reduce the side effects of immunosuppressants or to enhance immunity. Agarwood is the aromatic resinous portion of Aquilaria trees that has been traditionally used as a medicinal herb for the treatment of various diseases. Although previous studies have shown that agarwood can improve the body's immunity, evidence for this claim is still lacking. In this study, the immune-enhancing effects of the agarwood methanol extracts of Aquilaria malaccensis Lamk were evaluated in a RAW 264.7 macrophage model. Based on the results, the agarwood extracts markedly enhanced phagocytosis in the absence of cytotoxicity. The agarwood extract-treated RAW 264.7 cells exhibited the typical morphology of activated macrophages, which are spindle-shaped with elongated filopodia. Agarwood extract also significantly increased the production of nitric oxide (NO), which is associated with the increased expression of inducible NO synthase. Moreover, the secretion and expression levels of cytokines, such as tumor necrosis factor-α and interleukin (IL)-1β and IL-6, were increased by agarwood treatment. Notably, these are also associated with a mitogen-activated protein kinase signaling pathway. Taken together, our findings provide scientific evidence that agarwood has potential immune-enhancing effects in vitro.

• Journal of Life Science
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• v.31 no.5
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• pp.496-501
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• 2021

In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

• Journal of Life Science
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• v.32 no.7
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• pp.491-500
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• 2022

HK shiitake mushroom mycelium (HKSMM), containing 14% β-glucan, is a health functional food ingredient individually approved by the Korea Ministry of Food and Drug Safety for liver health. The anti-inflammatory effect of a 50% aqueous ethanol extract of HKSMM (designated HKSMM50) was studied in RAW 264.7 macrophage cells treated with lipopolysaccharide (LPS). An active hexose correlated compound (AHCC) was used as a positive control. LPS-activated RAW 264.7 cells were treated with HKSMM50 and AHCC (0, 20, 100, 500 ㎍/ml) and cultured for 24 hr. Inflammation-related elements in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA) kits, and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins in the cells was analyzed by Western blotting. The HKSMM50 lowered iNOS and COX-2 protein expressions, and nuclear factor-kappa B (NF-κB), nitric oxide (NO) and prostaglandin E₂ (PGE₂) contents in a concentration-dependent manner as compared to LPS treatment. Similarly, the HKSMM50 lowered the content of pro-inflammatory cytokines interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-6 (IL-6) contents and increased the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The efficacy of the AHCC treatment was similar to that of the HKSSM50 treatments. These results indicate that HKSMM50 showed an anti-inflammatory effect in LPS-treated RAW 264.7 cells by down-regulation of NF-κB signaling and suggest that HKSMM could be used as a health functional food ingredient to help improve immune function.