• Journal of Applied Biological Chemistry
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• v.66
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• pp.282-288
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• 2023

Due to the COVID-19 pandemic, the immuneenhancing health functional food market that protects our bodies from pathogens such as viruses continues to grow. In this study, we aimed to prove the Cheonggukjang, a high-nutrient food with high protein, fat, and dietary fiber content, as an immuneenhancing nutraceutical. Cheonggukjang water extract (CWE) increased the production of nitric oxide, reactive oxygen species, and cytokines such interleukin (IL)-6, IL-1β, and tumor necrosis factor-α without affecting viability in RAW 264.7 cells. Furthermore, CWE significantly upregulated the expression of inducible nitric oxide synthase and cyclooxygenase-2 in RAW 264.7 cells. CWE enhanced the phosphorylation of I kappa B kinase α/β and I kappa B (IκB)α, as well as the degradation of IκBα. CWE also induced increased phosphorylation of nuclear factor-kappa B p65 and facilitated the redistribution of p65 from the cytoplasm to the nucleus in RAW 264.7 cells. These findings suggest that CWE has potential as a health functional food material that can enhance the innate immune response.

• Biomolecules & Therapeutics
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• v.20 no.1
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• pp.50-56
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• 2012

WIN-34B showed analgesic and anti-inflammatory effects in various animal models of pain and osteoarthritis. However, the molecular mechanism by which WIN-34B inhibits pain and inflammation in vivo remains to be elucidated. We investigated the molecular mechanisms of the actions of WIN-34B using various in vitro models using fibroblast-like synoviocytes from patients with rheumatoid arthritis (RA FLSs), RAW264.7 cells and peritoneal macrophages. WIN-34B inhibited the level of IL-6, $PGE_2$, and MMP-13 in IL-$1{\beta}$-stimulated RA FLSs in a dose-dependent manner. The mRNA levels were also inhibited by WIN-34B. The level of $PGE_2$, NO, IL-$1{\beta}$, and TNF-${\alpha}$ were inhibited by WIN-34B at different concentrations in LPS-stimulated RAW264.7 cells. The production of NO and $PGE_2$ was inhibited by WIN-34B in a dose-dependent manner in LPS-stimulated peritoneal macrophages. All of these effects were comparable to the positive control, celecoxib or indomethacin. I${\kappa}B$B signaling pathways were inhibited by WIN-34B, and the migration of NF-${\kappa}B$ into the nucleus was inhibited, which is consistent with the degradation of $I{\kappa}B-{\alpha}$. Taken together, the results suggest that WIN-34B has potential as a therapeutic drug to reduce pain and inflammation by inhibiting the production of pro-inflammatory mediators.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.5
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• pp.537-544
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• 2017

In this study, the anti-inflammatory effect of Polyopes affinis ethanol extract (PAEE) was investigated using LPS-stimulated RAW 264.7 cells and a croton oil-induced ICR mice model. Treatment with PAEE significantly reduced production of nitric oxide (NO) and pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis factor $(TNF)-{\alpha}$, and $IL-1{\beta}$] in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. PAEE treatment also reduced expression of inducible NO synthase, cyclooxygenase-2, nuclear $factor-{\kappa}B$, and mitogen-activated protein kinases in LPS-stimulated RAW 264.7 cells. In the croton oil-induced ear edema test, application of PAEE (10~250 mg/kg body weight) reduced ear edema in a dose-dependent manner, and PAEE treatment at 50 mg/kg body weight showed similar inhibitory effects compared with prednisolone (10 mg/kg body weight). Histological analysis revealed reduced dermal thickness and lower number of infiltrated mast cells. These results suggest that PAEE might be used as a promising anti-inflammatory agent for inhibition of LPS-induced inflammation and ear edema formation.

• Biomolecules & Therapeutics
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• v.31 no.3
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• pp.330-339
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• 2023

Liver kinase B1 (LKB1) is a crucial tumor suppressor involved in various cellular processes, including embryonic development, tumor initiation and progression, cell adhesion, apoptosis, and metabolism. However, the precise mechanisms underlying its functions remain elusive. In this study, we demonstrate that LKB1 interacts directly with malic enzyme 3 (ME3) through the N-terminus of the enzyme and identified the binding regions necessary for this interaction. The binding activity was confirmed to promote the expression of ME3 in an LKB1-dependent manner and was also shown to induce apoptosis activity. Furthermore, LKB1 and ME3 overexpression upregulated the expression of tumour suppressor proteins (p53 and p21) and downregulated the expression of antiapoptotic proteins (nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and B-cell lymphoma 2 (Bcl-2)). Additionally, LKB1 and ME3 enhanced the transcription of p21 and p53 and inhibited the transcription of NF-κB. Moreover, LKB1 and ME3 suppressed the phosphorylation of various components of the phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B signaling pathway. Overall, these results suggest that LKB1 promotes pro-apoptotic activities by inducing ME3 expression.

• The Korea Journal of Herbology
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• v.30 no.5
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• pp.59-65
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• 2015

Objectives : Fermentation of herbs has been known to be helpful in improving the immune systems and protecting body against disease. The present study was conducted to evaluate anti-inflammatory effects of the fermentation extracts (FE) consisting of soybean, red ginseng andCitrus UnshiuPeel in lipopolysaccharide (LPS)-activated Raw264.7 cells.Methods : FE were prepared by the fermentation withBacillus Subtilisand then by extraction with ethanol (95%; prepared by the fermentation process). Cell viability was measured by MTT assay. Nitric oxide (NO) production was measured in culture media by Griess assay. The expression of nuclear factor (NF)-κB and inhibitory kappa B alpha (IκBα) was determined by Western blot.Results : LPS-induced production of NO and PGE₂was dose-dependently decreased by the treatment of FE in Raw264.7 cells. These suppressive effects of FE on NO and PGE₂production were related to the inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. FE inhibited LPS-induced production of pro-inflammatory cytokines, TNF-α, IL-6, and IL-1βin a dose-dependent manner. Furthermore, FE inhibited the NF-κB signaling pathway through the prevention of LPS-induced degradation of IκBαin cytosol and the nuclear translocation of NF-κB.Conclusions : These findings suggest that FE could have anti-inflammatory effects on LPS-induced inflammatory responses in macrophages.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.3
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• pp.991-994
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• 2012

Objective: VEGF-C has recently been identified as a key molecule which is involved in tumor lymphangiogenesis. The aim of this research was to investigate the role of PARP-1 inhibition in the regulation of VEGF-C expression in CT26 cells. Methods: CT26 cells were treated with or without the PARP-1 inhibitor 5-aminoisoquinolinone (5-AIQ). The expression of PARP-1, NF-kB, and VEGF-C proteins in CT26 cells was measured by Western blot analysis and the VEGF-C mRNA level was determined by reverse transcription polymerase chain reaction (RT-PCR). CT26-secreted VEGF-C was detected by enzyme-linked immunosorbent assay (ELISA). Results: The results of Western blot analysis showed that the expression levels of PARP-1, NF-kB, and VEGF-C were reduced in 5-AIQ treated CT26 cells and the levels of VEGF-C mRNA in 5-AIQ treated CT26 were significantly lower than t in 5-AIQ-untreated cells (P<0.05). The concentrations of CT26-secreted VEGF-C were also dramatically decreased (P<0.05). Conclusion: Here, we provide evidence for the first time that PARP-1 inhibition dramatically reduces VEGF-C expression via the nuclear factor NF-kB signaling pathway. We therefore propose that PARP-1 inhibition has an anti-lymphangiogenic effect and may contribute to the prevention of metastatic dissemination via the lymphatic system.

• Journal of Ginseng Research
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• v.45 no.5
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• pp.575-582
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• 2021

Background: In ginseng, there exists a glycolipoprotein complex with a special form of lipid LPAs called Gintonin. The purpose of this study is to show that Gintonin has a therapeutic effect on rheumatoid arthritis through LPA2 receptors. Methods: Fibroblast-like synoviocytes (FLS) were treated with Gintonin and stimulated with interleukin (IL)-1β. The antioxidant effect of Gintonin was measured using MitoSOX and H₂DCFDA experiments. The anti-arthritic efficacy of Gintonin was examined by analyzing the expression levels of inflammatory mediators, phosphorylation of mitogen-activated protein kinase (MAPK) pathways, and translocation of nuclear factor kappa B (NF-κB)/p65 into the nucleus through western blot. Next, after treatment with LPAR2 antagonist, western blot analysis was performed to measure inflammatory mediator expression levels, and NF-κB signaling pathway. Carrageenan/kaolin-induced arthritis rat model was used. Rats were orally administered with Gintonin (25, 50, and 100 mg/kg) every day for 6 days. The knee joint thickness, squeaking score, and weight distribution ratio (WDR) were measured as the behavioral parameters. After sacrifice, H&E staining was performed for histological analysis. Results: Gintonin significantly inhibited the expression of iNOS, TNF-α, IL-6 and COX-2. Gintonin prevented NF-κB/p65 from moving into the nucleus through the JNK and ERK MAPK phosphorylation in FLS cells. However, pretreatment with an LPA2 antagonist significantly reversed these effects of Gintonin. In the arthritis rat model, Gintonin suppressed all parameters that were measured. Conclusion: This study suggests that LPA2 receptor plays a key role in mediating the anti-arthritic effects of Gintonin by modulating inflammatory mediators, the MAPK and NF-κB signaling pathways.

• Nutrition Research and Practice
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• v.15 no.5
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• pp.591-603
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• 2021

BACKGROUND/OBJECTIVES: Unregulated inflammatory responses caused by hyperglycemia may induce diabetes complications. Hesperetin, a bioflavonoid, is a glycoside in citrus fruits and is known to have antioxidant and anticarcinogenic properties. However, the effect of inflammation on the diabetic environment has not been reported to date. In this study, we investigated the effect of hesperetin on proinflammatory cytokine secretion and its underlying mechanistic regulation in THP-1 macrophages with co-treatment LPS and hyperglycemic conditions. MATERIALS/METHODS: THP-1 cells differentiated by PMA (1 µM) were cultured for 48 h in the presence or absence of hesperetin under normoglycemic (5.5 mM/L glucose) or hyperglycemic (25 mM/L glucose) conditions and then treated with LPS (100 ng/mL) for 6 h before harvesting. Inflammation-related proteins and mRNA levels were evaluated by enzyme-linked immunosorbent assay, western blot, and quantitative polymerase chain reaction analyses. RESULTS: Hesperetin (0-100 µM, 48 h) treatment did not affect cell viability. The tumor necrosis factor-α and interleukin-6 levels increased in cells co-treated with LPS under hyperglycemic conditions compared to normoglycemic conditions, and these increases were decreased by hesperetin treatment. The TLR2/4 and MyD88 activity levels increased in cells co-treated with LPS under hyperglycemic conditions compared to normoglycemic conditions; however, hesperetin treatment inhibited the TLR2/4 and MyD88 activity increases. In addition, nuclear factor-κB (NF-κB) and Acetyl-NF-κB levels increased in response to treatment with LPS under hyperglycemic conditions compared to normoglycemic conditions, but those levels were decreased when treated with hesperetin. SIRT3 and SIRT6 expressions were increased by hesperetin treatment. CONCLUSIONS: Our results suggest that hesperetin may be a potential agent for suppressing inflammation in diabetes.

• Journal of the Korea Society of Computer and Information
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• v.27 no.1
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• pp.175-181
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• 2022

In this study, we tested the anti-inflammatory effects of broccoli leaf hexane fraction to confirm the applicability as a functional material in food and cosmetics. This sample was extracted using 70% ethanol from Broccoli leaf and then fractionated with hexane. The production of pro-inflammatory cytokines (TNF-α, IL-4, IL-6, IL-1β), protein expression of iNOS and COX-2, phosphorylation of MAPKs (ERK, JNK, p38) and NF-κB with broccoli leaf hexane fraction were assayed on LPS-stimulated RAW264.7 cells. The broccoli leaf hexane fraction inhibited the secretion of pro-inflammatory cytokines and protein expression of iNOS and COX-2. Also, the broccoli leaf hexane fraction reduced the phosphorylation of MAPKs and NF-κB. Therefore, it is considered that th broccoli leaf hexane fraction has the potential to be used as a natural anti-inflammatory material in food and cosmetics. In the future, it is considered necessary to study the anti-inflammatory mechanism and identification of major bioactive substances.

• Korean Journal of Organic Agriculture
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• v.29 no.1
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• pp.85-96
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• 2021

Reflux esophagitis (RE) is a common gastrointestinal disease observed at all ages, which seriously affects the quality of life. In this study, we investigated the anti-inflammatory effects of Pomacea canaliculata extract (PCE) using the experimental RE rat model. RE was induced by a surgical procedure. The rats were randomly divided into 4 groups: normal group, RE group, PCE group (RE treated with PCE, 100 mg/kg), positive control group (RE treated with ranitidine, 40 mg/kg). We performed the histological examination and measured the expression of tight junction complex and inflammatory mediators using western blot analysis. The phenotypes of RE were attenuated by PCE treatment. PCE administration significantly reduced esophageal mucosal damage and protected tight junction confirmed by claudin-5. Furthermore, PCE treatment reduced inflammatory reaction by inhibiting the expression of COX-2 and TNF-α. PCE treatment, also, reduced translocation of NF-κB into nuclear and IκB-α phosphorylation at the same time. Our findings indicate that PCE has the potential as a novel therapeutic agent to inhibit RE by protecting mucosal damage and regulating inflammatory reactions mediated by NF-κB signaling.