• 한방안이비인후피부과학회지
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• 제32권3호
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• pp.48-57
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• 2019

Objectives : This study examined the inhibitory effects of Wisaengtang(WST) on inflammatory mediators($NF-{\kappa}B$, COX-2, iNOS, IL-6) in cellular inflammatory responses induced by lipopolysaccharide(LPS). Methods : To investigate the cytotoxicity of WST, MTT assay was used. The inhibitory effects of inflammatory mediators were confirmed by real-time PCR and DPPH scavenging activity was measured to confirm the antioxidative effect. Results : When the $NF-{\kappa}B$ mRNA expression was inhibited, the levels of COX-2, iNOS, and IL-6 mRNA in the inflammatory response decreased significantly. iNOS is involved in the production of nitric oxide (NO), and it is confirmed that WST inhibits the expression of iNOS mRNA and thus the production of NO. Conclusions : These results suggest that WST can be a therapeutic substance for oxidation and inflammation through elimination of DPPH free radical and inhibition of $NF-{\kappa}B$ activity.

• 치위생과학회지
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• 제19권3호
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• pp.198-204
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• 2019

Background: Oxidative stress is a known to be associated with in the pathogenesis of many inflammatory diseases, including periodontitis. Ursolic acid is a pentacyclic triterpenoid with has antimicrobial, antioxidative, and anticancer properties. However, the role of ursolic acid in the regulating of osteogenesis remains undetermined. This study was aimed to elucidate the crucial osteogenic effects of ursolic acid and its ability to inhibit oxidative stress by targeting the immediate early response 3 (IER3)/nuclear factor erythroid 2-related factor 2 (Nrf2) pathway. Methods: Cell proliferation was determined using water-soluble tetrazolium salt assay, cell differentiation was evaluated by alkaline phosphatase (ALP) activity, and formation of calcium nodules was detected using alizarin red S stain. Generation of reactive oxygen species (ROS) was determined using by DCFH-DA fluorescence dye in hydrogen peroxide ($H_2O_2$)-treated MG-63 cells. Expression levels of IER3, Nrf2, and heme oxygenase-1 (HO-1) were analyzed using western blot analysis. Results: Our results showed that ursolic acid up-regulated the proliferation of osteoblasts without any cytotoxic effects, and promoted ALP activity and mineralization. $H_2O_2$-induced ROS generation was found to be significantly inhibited on treatment with ursolic acid. Furthermore, in $H_2O_2$-treated cells, the expression of the early response genes: IER3, Nrf2, and Nrf2-related phase II enzyme (HO-1) was enhanced in the presence of ursolic acid. Conclusion: The key findings of the present study elucidate the protective effects of ursolic acid against oxidative stress conditions in osteoblasts via the IER3/Nrf2 pathway. Thus, ursolic acid may be developed as a preventative and therapeutic agent for mineral homeostasis and inflammatory diseases caused due to oxidative injury.

• 융합정보논문지
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• 제11권1호
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• pp.236-243
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• 2021

본 연구에서는 개정향풀 열수추출물 (AW)과 메탄올추출물 (AM)의 기능성 화장품 활성을 조사하였다. DPPH 항산화 활성 결과에서 열수추출물은 90.5%의 높은 라디컬 소거 활성 (IC50 37.717±8.209 ㎍/mL)을 나타냈으며, ABTS 항산화 활성은 96.2%로 매우 높았으며, 이때 IC50은 185.244 ± 12.602 ㎍/mL이었다. 대식세포 RAW264.7에서 두 추출물은 LPS에 의한 NO, TNF-α, IL-6의 생성 및 iNOS 발현을 유의하게 억제하였다. 모든 추출물의 농도에서 세포독성은 거의 없었다. 수렴활성 결과 열수추출물이 74.33±2.48 mg/mL로 높은 수렴효능을 보였다. 따라서 이러한 결과들을 통하여 개정향풀 추출물은 항산화제, 항염 및 수렴제와 같은 미용 목적을 위한 천연제품, 식품첨가제 등 건강 증진에 기여할 것으로 기대된다.

• 한국응용과학기술학회지
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• 제38권1호
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• pp.309-317
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• 2021

본 연구는 침출차 원료로 레드비트를 활용하기 위해서 세포독성, 항염증 및 항산화 등의 활성을 조사하고자 설계되었다. 항산화 효능은 DPPH 및 ABTS 라디컬 소거 활성을 분석하여 평가하였다. RAW 264.7 세포를 통해 MTT 분석으로 세포 생존율을 평가하고 LPS로 유도하여 활성산소종과 염증매개체(일산화질소, TNF-α, IL-6 등)의 생성량을 확인하였다. 그 결과, DPPH 및 ABTS 라디컬 소거활성은 농도 의존적으로 증가하였으며, 모든 농도에서 세포독성이 없음을 확인하였다. 또한, LPS로 유도한 RAW 264.7 세포에서 활성산소종(ROS)과 일산화질소(NO), IL-6, TNF-α 생성량을 유의적으로 감소시켰다. 따라서 이러한 결과는 레드비트가 안전한 항산화 및 항염증 효과를 가진 침출차의 원료로서 높은 잠재력을 가지고 있음을 시사한다.

• Biomolecules & Therapeutics
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• 제30권6호
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• pp.540-545
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• 2022

Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

• 대한본초학회지
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• 제33권4호
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• pp.35-41
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• 2018

Objectives : Hyeonggaeyeongyotang Gagambang (HYT) is a herbal medicine prescribed for the treatment of inflammatory diseases, but it is necessary to study the exact therapeutic efficacy. This study aims to investigate the antibacterial and anti-inflmmatory activities of HYT. Methods : Antibacterial activity of HYT was confirmed by staining Escherichia coli, a gram negative strain, and Staphylococcus aureus, a gram positive strain, on solid Lysogeny Broth (LB) medium containing HYT. Antioxidant activity of HYT was confirmed by 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) assay. The phosphorylation of nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha ($I{\kappa}B{\alpha}$) after lipopolysaccharide (LPS) treatment with HYT-treated RAW 264.7 mouse macrophages cells was confirmed by immunoblot analysis and the level of interleukin 1 beta (IL-$1{\beta}$) mRNA expression level was confirmed by quantitative real-time PCR. Results : HYT showed a concentration-dependent antibacterial activity against Escherichia coli and Staphylococcus aureus and also showed excellent antioxidant activity. HYT treatment attenuated the phosphorylation of $I{\kappa}B{\alpha}$induced by LPS treatment in RAW 264.7 mouse macrophages cells. The phosphorylation of $I{\kappa}B{\alpha}$is crucial for the regulation of the expression of various pro-inflammatory cytokines. In addition, IL-$1{\beta}$ mRNA expression level of RAW 264.7 mouse macrophages cells stimulated by LPS treatment was also inhibited by HYT treatment. Conclusions : Through experimental demonstration of the antioxidative, antimicrobial and anti-inflammatory effects of HYT, we demonstrated that HYT is a herbal medicine effective for the treatment of inflammatory diseases caused by various bacterial infections.

• 대한통합의학회지
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• 제11권3호
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• pp.159-169
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• 2023

Purpose : Though other Citrus spp. have reported their anti-inflammatory and antioxidative activities in previous studies, the biological activity of Kiyomi (Citrus unshiu × C. sinensis) has not been reported yet. Therefore, this study attempted to analyze the anti-inflammatory mechanisms of Kiyomi leaf ethanol extract (KLEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : The cytotoxic effect of KLEE in RAW 264.7 cells was determined by WST-1 assay. Bacterial endotoxin, the concentration of nitric oxide (NO) was analyzed by the Griess reaction. In addition, Western blot analysis was applied to measure the protein expression level of inducible NO synthase (iNOS). The phosphorylated status of the critical inflammatory transcription factor, nuclear factor (NF)-𝜅B, and its upstream signaling molecules, phosphoinositide 3-kinase (PI3K)/Akt as well as mitogen-activated protein kinases (MAPKs), were also measured by Western blot analysis. Results : KLEE was not cytotoxic up to a concentration of 200 ㎍/㎖, and protein expression levels of iNOS and cyclooxygenase (COX)-2, enzymes that counteract NO and prostaglandin (PG) E2 production, were inhibited by KLEE treatment. The phosphorylated status of PI3K/Akt as well as MAPKs including extracellular regulated kinase (ERK), c-jun NH2kinase (JNK), and p38, were significantly attenuated by KLEE treatment in LPS stimulated RAW 264.7 cells. Moreover, one of phase II enzymes, heme oxygenase (HO)-1 which has known for its anti-inflammatory capacity, was strongly induced by KLEE treatment. Conclusion : Consequently, KLEE treatment significantly attenuated the production of NO as well as the expression levels of iNOS and COX-2 in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-𝜅B, as well as its upstream signaling molecules, PI3K/Akt and MAPKs, were also diminished by KLEE treatment with statistical significance in LPS-stimulated RAW 264.7 cells. These results suggest that KLEE might be a promising candidate for the attenuation of inflammatory disorders.

• Fisheries and Aquatic Sciences
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• 제26권2호
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• pp.158-168
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• 2023

The global output of Pyropia yezoensis (dried seaweed or laver, also called 'Gim' in Korea) has been reduced over the half-decade due to the wide spread of red rot disease, a serious algal disease affecting P. yezoensis. Recently, Gold 1 (G1), which is a resistant strain of P. yezoensis to red rot disease, was developed and commercialized in South Korea, yet its physiological activity has not been investigated. In this study, a comparative study was performed on G1 and commercially available strain of P. yezoensis (CP) for their antioxidative activities. Aqueous extract of G1 showed more marked 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity compared to that of CP. In 293T cells, antioxidant activity against H₂O₂-induced reactive oxygen species (ROS) formation was only observed in G1 extract. In addition, G1 extract showed more potent inhibitory effect on H₂O₂-induced apoptotic cell death than CP extract, as examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and fluorescence microscopy. Expression levels of various apoptosis-related genes, including B-cell lymphoma 2-associated X protein, p53, capase-3, and inflammatory cytokines, in H₂O₂-treated cells were significantly decreased by the treatment of G1. Taken together, the present study suggests that a new strain of red seaweed G1 can recover oxidative stress effectively by improving the imbalance of ROS generation and has a potential to be used a functional ingredient as an antioxidant source.

• 대한통합의학회지
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• 제11권2호
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• pp.169-179
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• 2023

Purpose : Although human periodontal ligament stem cells (hPDLSCs) are a supportive factor for tissue engineering, oxidative stress during cell culture and transplantation has been shown to affect stem cell viability and mortality, leading to failed regeneration. The aim of this study was to evaluate the antioxidant and protective effects against cell damage of celecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, and the antioxidant signal of hPDLSCs in H₂O₂-induced oxidative stress. Methods : To induce oxidative stress in cultured hPDLSCs, H₂O₂ was used as an exogenous reactive oxygen species (ROS). Dose-dependent celecoxib (.1, 1, 10, or 100 µM) was administered after H₂O₂ treatment. WST-1 assay was used to assess cell damage and western blot was used to observe antioxidant activity of hPDLSCs in oxidative stress. Immunohistochemistry was performed for inverting the localization of the SOD and Nrf2 antibody. Results : We found that progressive cell death was induced in hPDLSCs by H₂O₂ treatment. However, low-dose celecoxib reduced H₂O₂-induced cellular damage and eventually enhanced the SOD activity and Nrf2 signal of hPDLSCs. Oxidative stress-induced morphological change in hPDLSCs included lowered the survival and number of spindle-shaped cells, and shrinkage and shortening of cell fibers. Notably, celecoxib promoted cell survival function and activated antioxidants such as SOD and Nrf2 by positively regulating the cell survival signal pathway, and also reduced the number of morphological changes in hPDLS. Immunohistochemistry results showed a greater number of SOD- and Nrf2-stained cells in the celecoxib-treated group following oxidative stress. Conclusion : By increasing SOD and Nrf2 expression at the antioxidant system, the findings suggest that celecoxib enhanced the antioxidative ability of hPDLSCs and protected cell viability against H₂O₂-induced oxidative stress by increasing SOD and Nrf2 expression in the antioxidant system.

• 대한한방부인과학회지
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• 제22권1호
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• pp.53-78
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• 2009

Purpose: In this study, we investigated the anti-thrombotic and antiinflammatory efficacy of "Gamijoukyungtang(GJKT)". Methods: We studied inhibitory effects of platelet aggregation, FXa activation, $TXB_2$ and $PGE_2$ biosynthesis and suppressive effects of GPIIb/IIIa activity and oxidative damage, pro-inflammatory cytokine reduction effects of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)' in vitro. Also, we studied suppression of pulmonary embolism, AV shunt model in rats and shortening of Rat tail bleeding time in vivo. Results: GJKT/GJKT-1 extract showed inhibitory effects on GPIIb/IIIaactivities and platelet aggregation induced by ADP, epinephrine, collagen and arachidonic acid. They suppressed biosynthesis of $PGE_2$ but GJKT-1 only supressed biosynthesis of $TXB_2$. In FXa assay, they inhibited activation of FXa. they suppressed pulmonary embolism triggered by collagen and epinephrine. In AV shunt model, they decreased the weights of AV shunt thrombus. they inhibited pro-inflammatory cytokines and decreased oxidative damages caused by DPPH. Conclusion: We confirmed the anti-thrombosis, and ant-inflammatory efficacy of 'GJKT(by press extractor)/GJKT-1(by pressless extractor)'.