• BMB Reports
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• 제48권7호
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• pp.407-412
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• 2015

The 12 kDa FK506-binding protein (FK506BP12), an immunosuppressor, modulates T cell activation via calcineurin inhibition. In this study, we investigated the ability of PEP-1-FK506BP12, consisting of FK506BP12 fused to the protein transduction domain PEP-1 peptide, to suppress catabolic responses in primary human chondrocytes and in a mouse carrageenan-induced paw arthritis model. Western blotting and immunofluorescence analysis showed that PEP-1-FK506BP12 efficiently penetrated chondrocytes and cartilage explants. In interleukin-1β (IL-1β)-treated chondrocytes, PEP-1-FK506BP12 significantly suppressed the expression of catabolic enzymes, including matrix metalloproteinases (MMPs)-1, -3, and -13 in addition to cyclooxygenase-2, at both the mRNA and protein levels, whereas FK506BP12 alone did not. In addition, PEP-1-FK506BP12 decreased IL-1β-induced phosphorylation of the mitogen-activated protein kinase (MAPK) complex (p38, JNK, and ERK) and the inhibitor kappa B alpha. In the mouse model of carrageenan-induced paw arthritis, PEP-1-FK506BP12 suppressed both carrageenan-induced MMP-13 production and paw inflammation. PEP-1-FK506BP12 may have therapeutic potential in the alleviation of OA progression. [BMB Reports 2015; 48(7): 407-412]

• Journal of Veterinary Science
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• 제23권5호
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• pp.76.1-76.14
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• 2022

Background: Clinical dexamethasone (DEX) treatment or stress in bovines results in extensive physiological changes with prominent hyperglycemia and neutrophils dysfunction. Objectives: To elucidate the effects of DEX treatment in vivo on cellular energy status and the underlying mechanism in circulating neutrophils. Methods: We selected eight-month-old male bovines and injected DEX for 3 consecutive days (1 time/d). The levels of glucose, total protein (TP), total cholesterol (TC), and the proinflammatory cytokines interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α in blood were examined, and we then detected glycogen and adenosine triphosphate (ATP) content, phosphofructosekinase-1 (PFK1) and glucose-6-phosphate dehydrogenase (G6PDH) activity, glucose transporter (GLUT)1, GLUT4, sodium/glucose cotransporter (SGLT)1 and citrate synthase (CS) protein expression and autophagy levels in circulating neutrophils. Results: DEX injection markedly increased blood glucose, TP and TC levels, the Ca²⁺/P⁵⁺ ratio and the neutrophil/lymphocyte ratio and significantly decreased blood IL-1β, IL-6 and TNF-α levels. Particularly in neutrophils, DEX injection inhibited p65-NFκB activation and elevated glycogen and ATP contents and SGLT1, GLUT1 and GR expression while inhibiting PFK1 activity, enhancing G6PDH activity and CS expression and lowering cell autophagy levels. Conclusions: DEX induced neutrophils glucose uptake by enhancing SGLT1 and GLUT1 expression and the transformation of energy metabolism from glycolysis to pentose phosphate pathway (PPP)-tricarboxylic acid (TCA) cycle. This finding gives us a new perspective on deeper understanding of clinical anti-inflammatory effects of DEX on bovine.

• 동의생리병리학회지
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• 제31권4호
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• pp.220-225
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• 2017

Lithospermi Radix (LR) is known to have an anti-inflammatory effect. However, the mechanisms are not well known. In this study, LPS-induced mouse RAW 264.7 macrophage cells were treated with LR to investigate the time-dependent inflammation response of LR. RAW 264.7 cells were treated with various concentrations of LR for 24 hours, followed by MTS assay. Cell viability was increased at all experimental concentrations. The mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS were increased by treatment of RAW 264.7 cells with LR at a concentration of $200{\mu}g/ml$ for 6 hours and 24 hours. Treatment of LR with $200{\mu}g/ml$ concentration for 6 hours promoted mRNA expression levels of $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS in LPS-induced RAW 264.7 cells. However, $IL-1{\beta}$, $TNF-{\alpha}$ and iNOS mRNA expression was suppressed by treatment of LR with $200{\mu}g/ml$ concentration for 24 hours in LPS-induced RAW 264.7 cells. These results suggest that the effect on inflammation of LR is promptly promoted and then to rapidly alleviate the inflammatory reaction. This study proposes that the time-dependent activities of herbal medicine is a very important factor in analyzing the anti-inflammatory effect of various herbal medicines including LR.

• The Korean Journal of Physiology and Pharmacology
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• 제26권3호
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• pp.165-174
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• 2022

As the mechanism underlying glucose metabolism regulation by oxymatrine is unclear, this study investigated the effects of oxymatrine on pyroptosis in INS-1 cells. Flow cytometry was employed to examine cell pyroptosis and reactive oxygen species (ROS) production. Cell pyroptosis was also investigated via transmission electron microscopy and lactate dehydrogenase (LDH) release. Protein levels were detected using western blotting and interleukin (IL)-1β and IL-18 secretion by enzyme-linked immunosorbent assay. The caspase-1 activity and DNA-binding activity of nuclear factor kappa B (NF-κB) and nuclear factor (erythroid-derived 2)-like 2 protein (Nrf2) were also assessed. In the high glucose and high fat-treated INS-1 cells (HG + PA), the caspase-1 activity and LDH content, as well as Nod-like receptor family pyrin domain containing 3, Gsdmd-N, caspase-1, apoptosis-associated speck-like protein containing a CARD, IL-1β, and IL-18 levels were increased. Moreover, P65 protein levels increased in the nucleus but decreased in the cytoplasm. Oxymatrine attenuated these effects and suppressed high glucose and high fat-induced ROS production. The increased levels of nuclear Nrf2 and heme oxygenase-1 (HO-1) in the HG + PA cells were further elevated after oxymatrine treatment, whereas cytoplasmic Nrf2 and Keleh-like ECH-associated protein levels decreased. Additionally, the elevated transcriptional activity of p65 in HG + PA cells was reduced by oxymatrine, whereas that of Nrf2 increased. The results indicate that the inhibition of pyroptosis in INS-1 cells by oxymatrine, a key factor in its glucose metabolism regulation, involves the suppression of the NF-κB pathway and activation of the Nrf2/HO-1 pathway.

• 한국컴퓨터정보학회논문지
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• 제27권1호
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• pp.175-181
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• 2022

본 연구에서는 브로콜리 잎 헥산 분획물의 항염 효과를 평가하여 기능성 식품 및 화장품 소재로의 적용 가능성을 확인하였다. LPS-자극된 RAW264.7 세포에서 전염증성 사이토카인의 생성, iNOS와 COX-2의 발현, MAPK (ERK, JNK, p38) 및 브로콜리 잎 헥산 분획을 사용한 NF-κB의 인산화를 분석하였다. 브로콜리 잎 헥산 분획은 TNF-α, IL-4, IL-6, IL-1β 등의 전염증성 사이토카인의 분비와 iNOS와 COX-2의 발현을 억제했습니다. 또한, 브로콜리 잎 헥산 분획물은 MAPK와 NFκB의 인산화를 감소시켰다. 따라서 브로콜리 잎 헥산 분획물은 식품 및 화장품에서 천연 항염증 소재로 적용 가능성이 있는 것으로 판단된다. 향후 항염증 기전 및 주요 생리활성 물질의 규명에 대한 연구가 필요할 것으로 생각된다.

• 한방재활의학과학회지
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• 제31권1호
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• pp.33-46
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• 2021

Objectives The aim of this study is to evaluate anti-inflammatory and anti-arthritic effects of Sogyunghwalhyel-tang-gamibang (SGHHTGB) in cell and animal models and also to suggest one of putative mechanisms underlying its anti-arthritic effects. Methods Enzyme-linked immunosorbent assay was applied to measure the concentrations of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 and prostaglandin E2 (PGE2) in culture medium and blood serum and nitric oxide (NO) was assayed by Griess reagent. The expressions of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were analyzed by Western blot method. Results In a cell model using RAW264.7 macrophages stimulated with the endotoxin lipopolysaccharide (LPS), the drug, at its non-cytotoxic concentrations, inhibited the production of the pro-inflammatory cytokine TNF-α, IL-1β and IL-6. In addition, it suppressed the expression of the inflammatory enzyme iNOS and COX-2, and reduced the synthesis of the enzyme product NO (as stable nitrite) and PGE2 in activated macrophages. Meanwhile, in an animal model using rheumatic arthritis (RA) mice induced with injection of type II collagen antibody (CAb) and LPS, the drug improved clinical symptom of arthritis and reduced paw thickness and inflammatory cell infiltration. In blood of RA mice, the drug reduced serum levels of TNF-α, IL-1β, IL-6, nitrite, and PGE2, all inflammatory mediators produced by activated macrophages. Conclusions SGHHTGB may ameliorate CAb and LPS-induced RA in mice, presumably by inactivating macrophages that are capable of initiating joint inflammation by producing pro-inflammatory cytokines and expressing inflammatory enzymes.

• 동의생리병리학회지
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• 제18권6호
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• pp.1832-1842
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• 2004

The present study was performed to examine the possible anti-inflammatory effects of a herbal drug ASCH in RAW264.7 cell line. Inflammation was induced by LPS toxin treatment to RAW264.7 macrophage cell line. Increases in cytokine production such as IL-1β, IL-6. and IL-18, COX-2, NOS-Ⅱ (iNOS), and TNF-alpha were observed at mRNA level in the LPS-treated RAW264.7 cells. Measurement of IL-6, nitric oxide and the reactive oxygen species (ROS) showed increased production of these inflammation mediators. Treatment of ASCH effectively decreased IL-1β protein in a dose-dependent manner, and IL-6 and IL-18 were reduced at 100㎍/㎖ of ASCH concentration. NO production was also decreased by ASCH treatment. A slight inhibition for TNF-alpha in terms of protein, but not mRNA level was obtained by 100㎍/㎖ of ASCH treatment. ASCH treatment to normal RAW264.7 cells did not produce any cytotoxicity, indicting that the action of ASCH was selective to inflammatory cells. Thus, the present data suggest that ASCH may act as an important regulator to alleviate the inflammatory symptoms.

• 대한본초학회지
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• 제35권2호
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• pp.7-14
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• 2020

Objectives : Pyeongwi-san is widely used in Korean medicine for acute indigestion or gastrodynia. As a therapeutic agent for digestive diseases of modern people, in order to confirm the mechanism of Pyeongwi-san on digestive tract disease and the difference of therapeutic efficacy between its formulation, a comparative efficacy test was conducted on digestive tract disease animal model. Methods : For LPS enteritis animal model, male SD rats were intraorally treated with different formulation types of Pyeongwi-san, and then intraperitoneally administered LPS one hour later to induce enteritis. After 5 hours, blood was collected and TNFα, IL-1β, IL-6 and PGE2 were confirmed by ELISA. For acute gastritis animal model, male SD rats were intraorally treated with different formulation types of Pyeongwi-san according to the prescribed concentration, and then intraorally administered 60% ethanol and 150 mM HCl one hour later to induce acute gastritis. After 5 hours, blood was collected and TNFα,IL-6 were confirmed by ELISA. Results : In the LPS-administered enteritis animal model, Pyeongwi-san decreased TNFα, IL-1β, PGE2 and especially IL-6. Pyeongwi-san also decreased IL-6 in acute gastritis animal model. In addition, there was no significant difference in efficacy between the two formulations when compared with inflammatory markers. Conclusions : The efficacy of Pyeongwi-san was confirmed in the inflammatory markers related to digestive inflammatory diseases, and the efficay between two formulations of Pyeongwi-san was relatively similar. Further studies are needed to investigate the new applicability of Pyeongwi-san on different inflammatory diseases that have similar inflammation markers identified in this experiment.

• 동의생리병리학회지
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• 제18권1호
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• pp.114-121
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• 2004

We investigated the effects of healthful decoction utilizing Phellinus linteus(HDPL) for suppression in the process of carbon tetrachloride (CCl₄4)-induced inflammation(50% CCl₄ : olive oil=1:1, 1 ㎖/KgㆍB.W.) of rat using biochemical, Western, histological and immunohistochemical analysis. Biochemical analysis of serum showed that the level of aspartate aminotransferase, alanine aminotransferase, lactate Dehydrogenase, alkaline phosphatase and triglyceride were significantly decreased by pretreatment of HDPL, but albumin and nitric oxide were increased. Immunoblot analysis of the liver showed that CCl₄-induced expression of interleukin-1β(IL-1β) and inducible nitric oxide synthase(iNOS) was inhibited by pretreatment of HDPL. More severe histopathological changes of the liver such as Kupffer cell reaction, inflammatory cell infiltration and focal necrosis were demonstrated in the rats challenged with CCl₄ compared with normal. Fewer scores of these changes were observed in the HDPL pretreated rats. Immunohistochemical analysis of the liver showed that while the expression of IL-1β, iNOS, tumor necrosis factor-α, COX(cyclooxygenase)-1 and COX-2 tended to increase, a decline of these immunoreaction of HDPL pre-treated groups were observed in the hepatocytes, especially in the focal necrotic sites. These results suggest that HDPL may act as a therapeutic agent for liver disease through a regulation of inflammation-related proteins.

• Journal of Microbiology and Biotechnology
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• 제26권7호
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• pp.1198-1205
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• 2016

Lactic acid bacteria (LAB) have beneficial effects on intestinal health and skin diseases. Lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria, is known to induce the production of several cytokines such as TNF-α, IL-1β, and IL-8 and affect the intestinal microflora, anti-aging, sepsis, and cholesterol level. In this study, Weissella cibaria was isolated from Indian dairy products, and we examined its immune-enhancing effects. Live and heat-killed W. cibaria did not induce the secretion of immune-related cytokines, whereas LTA isolated from W. cibaria (cLTA) significantly increased the secretion of TNF-α, IL-1β, and IL-6 in a dose-dependent manner. cLTA increased the phosphorylation of nuclear factor kappa-light-chain-enhancer of activated B cells, p38 mitogen-activated protein kinases, and c-Jun N-terminal kinases in THP-1 cells. The secretion of TNF-α and IL-6 was also increased in the cLTA-treated mouse splenocytes. These results suggest that cLTA, but not W. cibaria whole cells, has immune-boosting potential and can be used to treat immunosuppression diseases.