• 대한한방부인과학회지
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• 제34권4호
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• pp.12-29
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• 2021

Objective: This study was performed to investigate the inhibitory effect of Cheongpochukeo-tang (CCT) on lipopolysaccharide (LPS)-induced inflammation model. Methods: RAW 264.7 cells were pre-treated with CCT and incubated with LPS (500 ng/ml) after 1 hour. Cell viability was measured by MTT assay to figure out cytotoxicity of CCT. The production of nitric oxide and mRNA expression of pro-inflammatory cytokine were measured. And the activation of mitogen-activated protein kinases (MAPKs) and nuclear factor kappa B (NF-κB) were examined to figure out molecular mechanisms of CCT's anti-inflammatory effects. In addition, mice survival rate and cytokine levels of serum were observed after treated with CCT. And mice liver tissues were observed and their cytokines levels in liver tissue were measured. Results: CCT did not have cytotoxic effect in RAW 264.7 cells. It inhibited LPS-induced nitric oxide (NO) production, but showed an increase in NO by itself at 2 mg/ml concentration. CCT inhibited mRNA expression of IL-1β, IL-6, TNF-α in a dose dependant and the activaton of MAPKs and NF-κB. In addition, CCT reduced mortality in the LPS-induced mouse model and inhibited production of cytokines in mouse serum and liver tissue. Conclusion: The results suggest that CCT could reduce LPS-induced inflammation by inhibiting MAPKs and NF-κB activaton, NO production, and pro-inflammatory cytokines secretion. Thereby, CCT could be effective medicine for the inflammatory disease.

• 대한본초학회지
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• 제36권1호
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• pp.67-76
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• 2021

Objective : Yukgunja-tang is one of the herbal prescriptions widely used for functional indigestion. In this study, we evaluated the pharmacological effect through the Yukgunja-tang formulation development. Methods : The RAW 264.7 cells were pretreated with Yukgunja-tang tablet (YGJT-T : 50, 100 and 200 ㎍/㎖) and then stimulated with lipopolysaccharide (LPS : 500 ng/㎖). Cell viability, inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) and inflammatory cytokines (IL-1β, IL-6, and TNF-α) were measured. Also, ICR mice induced acute gastritis by oral administration of 150 mM HCl in 60% ethanol. The YGJT-T (30 mg/kg) was pretreated for 3 days, and 150 mM HCl in 60% ethanol was orally administered 1 hour after the last drug treatment. Mice were sacrificed 1 hour after oral administration of 150 mM HCl in 60% ethanol. The gastric mucosa was observed, and inflammatory cytokines in the gastric tissue were measured. Results : The marker components of YGJT-T were determined by simultaneous analysis using HPLC. In RAW 264.7 cells, pretreatment of YGJT-T was non-toxic and inhibited the production of inflammatory mediators such as NO and PGE2 and suppressed inflammatory cytokines. In addition, pretreatment of YGJT-T improved bleeding and edema due to gastric lesions caused by acute gastritis and suppressed inflammatory cytokines. Conclusion : In summary, our results confirmed that treatment with YGJT-T has anti-inflammatory and anti-gastritis effects in vitro and in vivo. Therefore, in this study, YGJT-T could support a potential strategy for the prevention and treatment of gastritis.

• Journal of Microbiology and Biotechnology
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• 제30권12호
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• pp.1927-1936
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• 2020

Tunicates are known to contain biologically active materials and one species in particular, the sea peach (Halocynthia aurantium), has not been thoroughly studied. In this study we aimed to analyze the fatty acids profile of the H. aurantium body wall and its immunomodulatory effects on RAW264.7 macrophage-like cells. The fatty acids were classified into three categories: saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs). Omega-3 fatty acid content, including EPA and DHA, was higher than omega-6 fatty acids. H. aurantium body wall fatty acids exhibited enhanced immune response and anti-inflammatory effects on RAW264.7 macrophage-like cells. Under normal conditions, fatty acids significantly increase nitric oxide (NO) and PGE2 production in a dose-dependent manner, thereby improving the immune response. On the other hand, in LPS-treated RAW264.7 cells, fatty acids significantly decreased nitric oxide (NO) and PGE2 production in a dose-dependent manner, thereby enhancing anti-inflammatory effects. Fatty acids transcriptionally control the expression of the immune-associated genes, iNOS, IL-1β, IL-6, COX-2, and TNF-α, via the MAPK and NF-κB signaling cascades in RAW264.7 cells. However, in LPS-stimulated RAW264.7 cells, H. aurantium body wall fatty acids significantly inhibited expression of inflammatory cytokine; similarly, production of COX-2 and PGE2 was inhibited. The results of our present study provide insight into the immune-improving and anti-inflammatory effects of H. aurantium body wall fatty acids on macrophages. In addition, our study demonstrates that H. aurantium body wall is a potential source of immune regulatory components.

• 동의생리병리학회지
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• 제34권5호
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• pp.245-254
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• 2020

The purpose of this study was to investigate the effects of natural herb mixture containing Laminaria japonica Areschoung (LAM) on fine dust-induced bronchitis in mice. Laminaria japonica Areschoung is main content of LAM, which is including fucoidan. Fucoidan extracted from phaophyta is known to prevent bronchitis and to defend against bacteria and virus infection. In this study, we experienced the effect of LAM on bronchitis and investigated gene expressions (e.g ; IL-8, COX-2, MCP-1) and bio-markers (e.g ; IL-8, PGE2, MUC5AC) associated with bronchitis by using A549 cells. Also, we investigated whether LAM can suppress the bronchitis in fine dust-induced animal models. We injected fine dust (50 ㎕) twice as INT (Intra-Nasal-Trachea) method. Then LAM (200 mg/kg and 400 mg/kg) were oral administered for 14 days. We analyzed the number of immune cells, immunoglobulin E, bio-markers level associated with bronchitis. LAM significantly decreased bio-marker (IL-1β, IL-6, IL-8, TNF-α, Histamine, PGE2), immune cells (white blood cell, neutrophil, lymphocyte, monocyte), and immunoglobulin E, that are increased by fine dust. Taken together, this study suggest that LAM can be used as effective herbal extract for bronchitis.

• 한국식품영양학회지
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• 제34권1호
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• pp.36-46
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• 2021

After ethanol (BM-E and RW-E) and hot-water (BM-HW and RW-HW) extracts were fractionated from two herbal mixtures (BM and RW), their physiological activities were investigated. All extracts consisted of more than 50% of neutral sugar, with their total polyphenol levels higher than flavonoid levels. Radical scavenging activities of EtOH extracts remained significantly higher compared to that of hot-water extracts, and in particular, RW-E showed consistently higher antioxidant activity than BM-E. When anti-inflammatory activities of the extracts were evaluated by LPS-stimulated RAW 264.7 cells at 10~500 μg/mL non-cytotoxicity doses, BM-E showed significantly higher levels of TNF-α, IL-1β, IL-6, and nitric oxide inhibitory activity than those of hot-water extracts and RW-E. Murine peritoneal macrophage cells were shown to be enhanced in crude polysaccharides (BM-CP and RW-CP fractionated from BM-HW and RW-HW) compared to hot-water extracts and polysaccharide K (PSK, positive control). Especially, RW-CP exhibited higher activity than BM-CP, and component sugar analysis showed that BM-CP mainly contained galacturonic acid, glucose, arabinose, galactose, and xylose (34.5%, 33.9%, 16.1%, 7.1%, and 6.3%, respectively), whereas RW-CP showed different measurements (29.5%, 59.2%, 5.0%, 4.5%, and 0.2%). In conclusion, two herbal mixtures could contain varying sets of physiological activities dependent on different extraction and fractionation methods.

• Biomolecules & Therapeutics
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• 제29권5호
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• pp.536-544
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• 2021

5-amino salicylic acid (5-ASA) is a standard therapy for the treatment of mild to moderate forms of inflammatory bowel diseases (IBD) whereas more severe forms involve the use of steroids and immunosuppressive drugs. Hyaluronic acid (HA) is a naturally occurring non-sulfated glycosaminoglycan that has shown epithelium protective effects in experimental colitis recently. In this study, both 5-ASA (30 mg/kg) and HA (15 mg/kg or 30 mg/kg) were administered rectally and investigated for their potential complementary therapeutic effects in moderate or severe murine colitis models. Intrarectal treatment of moderate and severe colitis with 5-ASA alone or HA alone at a dose of 30 mg/kg led to a significant decrease in clinical activity and histology scores, myeloperoxidase activity (MPO), TNF-α, IL-6 and IL-1β in colitis mice compared to untreated animals. The combination of HA (30 mg/kg) and 5-ASA in severe colitis led to a significant improvement of colitis compared to 5-ASA alone. Combined rectal therapy with HA and 5-ASA could be a treatment alternative for severe cases of IBD as it was the only treatment tested that was not significantly different from the healthy control group. This study further underlines the benefit of searching for yet unexplored drug combinations that show therapeutic potential in IBD without the need of designing completely new drug entities.

• 한국미생물·생명공학회지
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• 제50권1호
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• pp.22-30
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• 2022

Haliotis discus hannai called abalone, is the valuable marine mollusks and the by-products of abalone processing are viscera. Brownish abalone male viscera (AMV), which have not been reported as having anti-inflammatory effects, was extracted with acetone and fractionated by different six acetone/hexane ratios (0, 10, 20, 30, 40, and 100%) using a silica column via in vitro ABTS and DPPH radical and nitric oxide (NO) production assay-guided fractionation. Among the fractions, the acetone/hexane ratio 40%, A40 exhibited the most potent radical scavenging activities and inhibition of lipopolysaccharide (LPS)-induced NO production without cytotoxicity. A40 inhibited LPS-induced intracellular reactive oxygen species (ROS) production in a dose-dependent manner. Western blot analysis revealed that A40 down-regulated the activation of NF-κB, MAPK (ERK 1/2, p-38, and JNK), and inflammatory enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2. Moreover, this fraction inhibited the generation of pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. These results suggested that AMV containing A40 with anti-inflammatory and anti-oxidantive effects, is the effective therapeutic and functional material for treating inflammatory disorders.

• Journal of Microbiology and Biotechnology
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• 제32권5호
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• pp.638-644
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• 2022

Probiotics modulate the gut microbiota, which in turn regulate immune responses to maintain balanced immune homeostasis in the host. However, it is unclear how probiotic bacteria regulate immune responses. In this study we investigated the immunomodulatory effects of heat-killed probiotics, including Lactiplantibacillus plantarum KC3 (LP3), Lactiplantibacillus plantarum CKDB008 (LP8), and Limosilactobacillus fermentum SRK414 (LF4), via phagocytosis, nitric oxide (NO), and pro-inflammatory cytokine production in macrophages. We thus found that heat-killed LP8 could promote the clearance of foreign pathogens by enhancing the phagocytosis of macrophages. Treatment with heat-killed LP8 induced the production of NO and pro-inflammatory cytokines, including TNF-α, IL-6, and IL-1β. In addition, heat-killed LP8 suppressed the production of NO and cytokines in LPS-induced RAW264.7 cells, suggesting that heat-killed LP8 exerts immunomodulatory effects depending on the host condition. In sum, these results indicate that heat-killed LP8 possesses the potential for immune modulation while providing a molecular basis for the development of functional probiotics prepared from inactivated bacterial cells.

• Journal of Microbiology and Biotechnology
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• 제32권7호
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• pp.885-891
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• 2022

Plant-based protein sources such as soybean meal have low digestibility and are generally promoted accumulation of undigested proteins into the intestine by enzymatic treatments. Moreover, potential intestinal pathogens ferment undigested proteins, producing harmful substances, such as ammonia, amines and phenols, leading to an overactive immune response and diarrhea in weaned pigs. As a solution, dietary proteases hydrolyze soybean-based antinutritive factors, which negatively affect immune responses and gut microbiota. In this study, we investigated the effects of dietary proteases (PRO) in a low-crude protein (CP) commercial diet on the immune responses and gut microbiota of weaned pigs. The experimental design consisted of three dietary treatments: a commercial diet as a positive control (PC; phase1 CP = 23.71%; phase 2 CP: 22.36%), a lower CP diet than PC as negative control (NC; 0.61% less CP than PC), and NC diet supplement with 0.02% PRO. We found that PRO tended to decrease the frequency of diarrhea in the first two weeks after weaning compared with PC and NC. In addition, pigs fed PRO showed decreased TNF-α and TGF-β1 levels compared with those fed PC and NC. The PRO group had a higher relative proportion of the genus Lactobacillus and lower levels of the genus Streptococcus than the PC and NC groups. In conclusion, the addition of PRO to a low CP commercial weaned diet attenuated inflammatory responses and modified gut microbiota in weaned pigs.

• Journal of Microbiology and Biotechnology
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• 제32권9호
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• pp.1146-1153
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• 2022

Many probiotic species have been used as a fermentation starter for manufacturing functional food materials. We have isolated Bifidobacterium animalis subsp. lactis LDTM 8102 from the feces of infants as a novel strain for fermentation. While Glycine max has been known to display various bioactivities including anti-oxidant, anti-skin aging, and anti-cancer effects, the immune-modulatory effect of Glycine max has not been reported. In the current study, we have discovered that the extract of Glycine max fermented with B. animalis subsp. lactis LDTM 8102 (GFB 8102), could exert immuno-modulatory properties. GFB 8102 treatment increased the production of immune-stimulatory cytokines in RAW264.7 macrophages without any noticeable cytotoxicity. Analysis of the molecular mechanism revealed that GFB 8102 could upregulate MAPK2K and MAPK signaling pathways including ERK, p38, and JNK. GFB 8102 also increased the proliferation rate of splenocytes isolated from mice. In an animal study, administration of GFB 8102 partially recovered cyclophosphamide-mediated reduction in thymus and spleen weight. Moreover, splenocytes from the GFB 8102-treated group exhibited increased TNF-α, IL-6, and IL-1β production. Based on these findings, GFB 8102 could be a promising functional food material for enhancing immune function.