• The Korea Journal of Herbology
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• v.31 no.1
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• pp.69-75
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• 2016

Objectives : The aim of this study is to investigate the various effects of individual or combined extract of GamiSaengmaeksan (GSS) on cell viability, anti-inflammatory and antioxidant activityMethods : In order to evaluate cytotoxicity, MTT assay was performed. We investigated the levels of proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 and interleukin (IL)-1β, and nitric oxide(NO) in LPS-induced RAW 264.7 cells to check the effects on anti-inflammatory activity. The level of NO production in RAW 264.7 cells was measured by using Griess reagent. The levels of cytokines and ROS were measured by Luminex and Flow cytometry, respectively.Results : At concentration of 200 ㎍/㎖ GSS, cytotoxicity was observed in RAW 264.7 cells. However, at concentration less than 100 ㎍/㎖ of both combine and individual GSS, cytotoxicity was not observed in Raw 264.7 cells. However, the level of ROS in RAW 264.7 cells were decreased at both extract of 100 ㎍/㎖ GSS. Also, the level of NO in RAW 264.7 cells were decreased from extraction of concentration of 100 ug/ml in GSS and individual-extraction of Liriopis Tuber, White Ginseng and Glycyrrhizae Radix. In addition, productions of pro-inflammatory cytokines (TNF-α) in LPS-induced RAW 264.7 cells were decreased from extraction of concentration of 10 and 100 (㎍/㎖) in GSS and individual-extraction of Liriopis Tuber.Conclusions : It is concluded that combined extract of GSS appears to be more effective in anti-oxidation and anti-inflammatory effect than those in individual-extraction of GSS. These results may be developed as a raw material for new therapeutics to ease the symptoms related with inflammatory and oxidative stress.

• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.309-317
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• 2021

This study was designed to examine the cell cytotoxic, anti-inflammatory, and antioxidant activity for raw material of Beta vulagaris tea. Antioxidative ability was evaluated by bioassays using DPPH and ABTS radical scavenging activity. Cell viability was assessed by MTT assay using RAW 264.7 cells, and investigated production levels of reactive oxide speies, and inflammatory meditors(i.e., nitric oxide, tumor necrosis factor-α, and interleukin-6) in LPS-induced RAW 264.7 cells. As a results, DPPH and ABTS raidcal scavenging activity were increased in a dose-dependent manner, and confirmed no cytotoxicity in all concentration. Also, it was significantly decreased level of ROS, NO, IL-6, and TNF-a in LPS-induced RAW 264.7 cells. Therefore these results suggest Beta vulagaris has considerable potential as a raw material of leached tea with safe anti-oxidative and anti-inflammatory effects.

• Journal of Life Science
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• v.31 no.12
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• pp.1110-1119
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• 2021

The purpose of this study was to investigate whether the inflammatory response in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages could be promoted by particulate matter 2.5 (PM_2.5) stimulation. To this end, the levels of inflammatory parameters, reactive oxygen species (ROS) and inflammation-regulating genes were investigated in RAW 264.7 cells treated with PM_2.5 in the presence or absence of LPS. Our results showed that the production levels of pro-inflammatory mediators (nitric oxide and prostaglandin E₂) and cytokines (interleukin-6 and -1β) were significantly increased by PM_2.5 stimulation in LPS-treated RAW 264.7 cells, which was correlated with increased expression genes involved in their production. In addition, when LPS-treated RAW 264.7 cells were exposed to PM_2.5, nuclear factor-kappaB (NF-κB) expression was further increased in the nucleus, and the expression of inhibitor of NF-κB as well as NF-κB in the cytoplasm was decreased. These results suggest that the co-treatment of PM_2.5 and LPS further increases the activation of the NF-κB signaling pathway compared to each treatment alone, thereby contributing to the promotion of transcriptional activity of inflammatory genes. Furthermore, although the generation of ROS was greatly increased by PM_2.5 in LPS-treated RAW 264.7 cells, the NF-κB inhibitor did not reduce the generation of ROS. In addition, when the generation of ROS was artificially suppressed, the production of inflammatory mediators and the activation of NF-κB were both abolished. Therefore, our results suggest that the increase in the NF-κB-mediated inflammatory response induced by PM_2.5 in LPS-treated RAW 264.7 macrophages was a ROS generation-dependent phenomenon.

• Journal of Oriental Neuropsychiatry
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• v.22 no.2
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• pp.107-128
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• 2011

Objectives : This experiment was designed to investigate the efficacy of DDTCMT hot water extract & ultra-fine powder on Alzheimer's Disease Model. Methods : The effects of the DDTCMT hot water extract on expression of IL-$1{\beta}$, IL-6, TNF-${\alpha}$, COX-2, NOS-II, IL-10, IL-1 receptor antagonist mRNA and production of IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by lipopolysacchaide(LPS) were investigated. Expression of NO, ROS in BV2 microglial cell line treated by LPS and AChE activity in PC-12 cell treated by NGF were investigated. anti-AChE was observed through Western blot analysis. The effects of the DDTCMT hot water extract & ultra-fine powder on the behavior of the memory deficit mice induced by scopolamine were investigated. Results : 1. The DDTCMT hot water extract significantly decreased the production of mIL-6, mNOS-II, mTNF-${\alpha}$, and increased the production of mIL-10, mIL-1 receptor antagonist. 2. The DDTCMT hot water extract significantly suppressed the production of IL-$1{\beta}$, IL-6, TNF-${\alpha}$ in BV2 microglial cell line treated by LPS. 3. The DDTCMT hot water extract significantly suppressed the NO and ROS production in BV2 microglial cell line treated by LPS. 4. The DDTCMT hot water extract groups showed inhibition of AChE activity in NGF treated PC-12 cell line. 5. The DDTCMT hot water extract suppressed anti-AChE expression in NGF treated PC-12 cell line was observed by Western blot analysis. 6. The DDTCMT hot water extract & ultra-fine powder groups showed significantly inhibitory effect on the scopolamine -induced impairment of memory in the experiment of Morris water maze. Conclusions : These results suggest that the DDTCMT hot water extract & ultra-fine powder may be effective for the prevention and treatment of Alzheimer's disease.

• The Korea Journal of Herbology
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• v.36 no.5
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• pp.125-133
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• 2021

Objectives : Thymus quinquecistatus var. japonica (H.Hara) is a member of the genus Thymus of perennial aromatic herb, and it's designated as a natural monument of South Korea. It has traditionally been known to have protective or therapeutic effects on various human disease including cerebrovascular and neurological disease. Recently it was suggested that essential oil extracted from thyme has anti-fungal and anti-bacterial effect. The aim of this study is to investigate anti-inflammatory effect of Thymus quinquecistatus var. japonica in Raw 264.7 macrophage cell line. Methods : The cytotoxic effects of water and 70% ethanol extracts from Thymus quinquecistatus var. japonica, was tested using MTT assay. Inhibitory effects of the extracts to nitric oxide production and mRNA expression of inflammatory cytokines were examined by RT-PCR. Also, MitoSOX-red assay and JC-1 assay were performed to determine if the extracts can inhibit mitochondrial ROS accumulation and maintain mitochondrial membrane potential. Results : In LPS-induced inflammatory response, the extracts efficiently reduced nitric oxide NO production through inhibiting mRNA expression of iNOS enzyme. In addition, expression of the proinflammatory cytokines, IL-1𝛽 and IL-6, was also down-regulated by the extract treatments. Excessive accumulation of mitochondrial ROS induced by LPS was inhibited in the extract treated cells, which finally protected mitochondrial membrane potential. Conclusions : These results showed that water and 70% ethanol extracts from Thymus quinquecistatus var. japonica have anti-inflammatory effect through down regulation of IL-1𝛽, IL-6, and iNOS, and also have antioxidative effect against mitochondrial ROS accumulation that promote inflammatory response.

• Natural Product Sciences
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• v.23 no.3
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• pp.162-168
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• 2017

Cordyceps bassiana has long been used as an oriental medicine and reported to possess diverse biological activities. The fruiting bodies of Cordyceps bassiana was extracted with ethanol and then further fractionated with n-hexane, ethyl acetate, n-butanol and water. The butanol fraction from Cordyceps bassiana (CBBF) exhibited the most effective in anti-inflammatory activity in RAW 264.7 macrophages and the roles of CBBF on the anti-inflammation cascade in LPS-stimulated RAW 264.7 cells were studied. To investigate the mechanism by which CBBF inhibits NO, iNOS and COX-2, the activation of $I{\kappa}B$ and MAPKs in LPS-activated macrophage were examined. Our present results demonstrated that CBBF inhibits NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cells, and these effects were mediated through the inhibition of $I{\kappa}B-{\alpha}$, JNK and p38 phosphorylation. Also, CBBF suppressed activation of MAPKs including p38 and SAPK/JNK. Furthermore, CBBF significantly suppressed LPS-induced intracellular ROS generation. Its inhibition on iNOS expression, together with its antioxidant activity, may support its anti-inflammatory activity. Thus Cordyceps bassiana can be used as a useful medicinal food or drug for further studies.

• Journal of Marine Bioscience and Biotechnology
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• v.11 no.2
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• pp.71-80
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• 2019

Previously, agar obtained from Gelidum sp. has a small molecular weight and has the disadvantage of inherent viscosity properties and poor functionality as a dietary fiber. In order to improve aforementioned disadvantages, agar having a fluidity that can be added to food at a higher concentration that a powder agar having a gelling property at low concertation was manufactured. In addition, the anti-inflammatory activity of agar hydrolysates was evaluated to confirm their potential as a functional material. As a result, agar hydrolysates significantly reduced NO levels secreted by LPS-activated macrophages and inhibited the expression of iNOS and COX-2, which are inflammatory mediators that regulates NO secretion in macrophages. Furthermore, in in vivo zebrafish embryos model results demonstrated significant reduction of LPS induced NO production after the treatment of agar hydrolysate hydrolyzed for 360 min. In addition, ROS production and cell death by stresses were also reduced in LPS-exposed embryos after the treatment of agar hydrolysis product hydrolyzed for 360 min. Taken together, agar hydrolysate hydrolyzed for 360 min can be easily added into food due to their fluidity and used as a food ingredient that inhibits inflammation due to their anti-inflammatory property.

• The Journal of Internal Korean Medicine
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• v.26 no.1
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• pp.107-118
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• 2005

Objectives : Peroxynitrite $(ONOO^-)$, fonned from the reaction of $O_2^-$ and NO, is a cytotoxic species that can oxidize several cellular components such as proteins, lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. Due to the lack of endogenous enzymes to thwart $ONOO^-$ activation, developing a specific $ONOO^-$ scavenger is remarkably important. The aim of this study was to investigate scavenging activities of $ONOO^-$ and its precursors, NO and $O_2^-$ and its scavenging mechanism of Ojawhan. Methods : To investigate scavenging activities of $ONOO^-$, NO, $O_2^-$ and its scavenging mechanism using fluorescent probes, DCFDA, DAF-2 and DHR 123. The $ONOO^-$ scavenging activity on Ojawhan was assayed by measuring oxidized dihydrorhodamine 123 (DHR 123) by fluorometry. Oxidative stress was induced by strong oxidants t-butyl hydroperoxide (t-BHP). Endothelial cell (YPEN-1) was used for detection of intracellular oxidative stress. Results : Ojawhan markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthennore, ${\underline{Ojawhan}}$ blocked lipopolysaccharide (LPS)-induced $ONOO^-$, $O_2^-$ and NO generation utilizing kidney homogenates of LPS-injected mouse and inhibited t-BHP-induced ROS and $ONOO^-$ in endothelial cell culture system. Conclusions : These results suggest that Ojawhan be developed as an effective $ONOO^-$ scavenger for the prevention of $ONOO^-$ involved diseases and age-related diseases.

• Natural Product Sciences
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• v.24 no.1
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• pp.28-35
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• 2018

Pulegone is a naturally occurring organic compound obtained from essential oils from a variety of plants. The aim of this study was to investigate the anti-inflammatory effects through the inhibitory mechanism of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), nuclear factor kappa B ($NF-{\kappa}B$), mitogen-activated protein kinases (MAPK) pathways and the activation of nuclear factor erythroid 2-related factor 2 (Nrf2)/ heme oxygenase (HO)-1 pathways in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Results revealed that pulegone significantly inhibited NO production as well as iNOS and COX-2 expressions. Meanwhile, western blot analysis showed that pulegone down-regulated LPS-induced $NF-{\kappa}B$ and MAPKs activation in RAW 264.7 cells. Furthermore, the selected compound suppressed LPS-induced intracellular ROS production in RAW 264.7 cells, while the expression of stress response gene, HO-1, and its transcriptional activator, Nrf-2 was upregulated upon pulegone treatment. Taking together, these findings provided that pulegone inhibited the LPS-induced expression of inflammatory mediators via the down-regulation iNOS, COX-2, $NF-{\kappa}B$, and MAPKs signaling pathways as well as up-regulation of Nrf-2/HO-1 indicating that pulegone has a potential therapeutic and preventive application in various inflammatory diseases.

• The Journal of Pediatrics of Korean Medicine
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• v.28 no.3
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• pp.47-58
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• 2014

Objectives GyejigaChulBuTang (GCBT) is a prescription used to treat acute and chronic arthritis in Korea, China, and Japan. This study assessed the anti-inflammatory and anti-oxidant activities of GCBT on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Methods Raw264.7 cells were pretreated with or without GCBT for 1 hour prior to incubation with LPS. Anti-inflammatory activity of GCBT was evaluated with reference to gene expression and production levels of proinflammatory cytokines ($TNF{\alpha}$, IL-$1{\beta}$, IL-6, GM-CSF and $INF{\gamma}$) and inflammatory mediators (iNOS, COX-2, NO and $PGE_2$). In addition, intracellular ROS generation and signal transduction of MAPK family, PI3K/Akt and $I{\kappa}B{\alpha}/NF{\kappa}B$ was investigated. Results Prior treatment with GCBT inhibited elevation of $TNF{\alpha}$, IL-$1{\beta}$, IL-6, GM-CSF, $INF{\gamma}$, NO and $PGE_2$, together with their cognate mRNAs in a dose-dependent manner. Intracellular ROS contents were similarly reduced. These effects were due to inhibition of LPS-induced phosphorylation of MAPK family, PI3K/Akt and $I{\kappa}B{\alpha}$ as well as nuclear translocation of $NF{\kappa}B$. Conclusions GCBT suppresses pro-inflammatory mediators. GCBT has potential in the treatment of juvenile rheumatoid arthritis associated with inflammation.