Background : As a part of ongoing research to elucidate and characterize anti-inflammatory nutraceuticals, six kinds of plant extracts (aerial part of Nepeta cataria, leaves of Lonicera maackii, leaves of Platycarya strobilacea, flower of Fagopyrum dibotrys, flowers and fruits of Solanum nigrum, stem of Physostegia virginiana) were tested for their ability to suppress inflammation. The anti-inflammatory has been studied in lipopolysaccharide (LPS)-stimulated RAW264.7 cells which cells synthesized nitric oxide (NO) from L-arginine by nitric oxide synthase (NOS). In this study, NO synthesis inhibitory activity of six kinds of plant extracts on LPS-stimulated RAW 264.7 mouse macrophages was evaluated. Methods and Results : Six kinds of plant extracts were parceled out from RDA (Rural Development Administration). RAW 264.7 cells (1.5×105 cells/well) were seeded onto 96-well plates with DMEM media containing 10% FBS and 1% antibiotics. The cells were pretreated with the extracts and LPS-stimulated cells for 24 h. Cellular NO production was stimulated by adding 1 μg/mL of LPS. After incubation, Griess reagent was used to determine NO production. Absorbance was measured at 520 nm by microplate reader. NO synthesis inhibitory activity potential of these extracts was evaluated by assessing NO production by LPS-stimulated RAW 264.7 cells in the presence. As a result, inhibition rate of NO production was about 40% of L. maackii, 33% of F. dibotrys, 23% of P. strobilacea and 17% of P. virginiana. Meanwhile, there was no significant results in aerial part of N. cataria and flowers and fruits of S. nigrum. Conclusion : From the above results, we be able to confirm that leaves of L. maackii and flower of F. dibotrys appeared dose-dependent NO synthesis inhibitory activity and leaves of P. strobilacea appeared NO synthesis inhibitory activity in low-concentration. As screening NO synthesis inhibition of six extracts, they may be a good candidate for delaying the progression of human inflammatory diseases and warrants further studies.

Background : Geranium Koreanum(GK), a species of the family Geranium is a perennial herb plants. We performed to determine the anti-inflammatory effects in Lipopoly -saccaride induced RAW 264.7 cell of ethanol extracts from Geranium koreanum. Methods and Results : The dried whole plants 50g of G. koreanum was refluxed three times in 0.5 L 70% ethanol for 2hr. The RAW 264.7 cells for anti-inflammatory assay were obtained from the American Tissue Culture Collection(Manassas, VA), and it cultured in DMEM containing 10% Fetal Bovine Serum(FBS) in 5%, CO2 incubator. An anti-inflammatory of G. koreanum measured by Ntrite oxide(NO) production and the protein expression levels of pro-inflammatory proteins such as COX-2 and iNOS, reductions in activation of NF-kB transcription factor. The results showed that G. koreanum was inhibited NO production and exhibited nontoxic in concentration 50~200㎍/㎖. Also, G. koreanum extracts indicate a significant reduction activation of NF-kB transcription factor and inhibition of pro-inflammatory proteins such as COX-2 and iNOS. Conclusion : The above results suggested that G. koreanum extracts expected the antiinflammatory effects and development possibility as nutraceuticals.

Background : Achyranthes japonica Nakai (AJ) is a perennial herb with a wide distribution in East Asia including Korea, China, and Japan, and it is mainly used as a medicinal plant. In Korea, AJ has been widely used to control pain and improve symptoms in OA patients. AJ contains several important phytochemicals such as saponins, inokosterone, ecdysterone, and oleanolic acid bisdesmoside. Methods and Results : The aim of this work was to investigate the antioxidant and anti-inflammatory activities of fermented and ethanol extracts of Achyranthes japonica Nakai (AJ). The extracts showed strong reductive power and nitrite scavenging, hydroxyl radical scavenging, superoxide radical scavenging, and DNA damage prevention activities. Treatment of RAW 264.7 macrophages with AJ inhibited lipopolysaccharide (LPS)-induced NO secretion and iNOS expression without affecting cell viability. AJ also inhibited cyclooxygenase 2 (COX-2) expression, leading to the suppression of COX-2-derived prostaglandin E2 production. These inhibitory effects of AJ were accompanied by reduced production of tumor necrosis factor-α and interleukins (IL)-1β, -6, and -10. Furthermore, AJ suppressed LPS-induced phosphorylation of extracellular signal regulated kinase, c-Jun N-terminal kinase, and p38. Moreover, AJ inhibited malondialdehyde production and myeloperoxidase activity in LPS-stimulated RAW 264.7 cells. Conclusion : The antioxidant activity of plants is closely related to their medicinal properties and is widely used as a parameter to determine the bioavailability of medicinal plants. The antioxidant and biological activities of AJ extracts might be due to the synergistic actions of multiple bioactive compounds. It can be concluded that AJ extracts are a potential source of biologically important drug candidates.

Background : Ganoderma lucidum is a non-toxic, medicinal mushroom, which is known to possess anti-inflammatory and immunomodulating activities. However, the effects and mechanism of action of Ganoderma lucidum on lipopolysaccharide (LPS) and interferon-gamma (IFN-γ)-induced nitric oxide (NO) production and its-related cytokine expression are not yet fully understood. This study aimed to evaluate the effect of Ganoderma lucidum on NO production and NO-mediated pro-inflammatory cytokine expression in LPS/IFN-γ-induced RAW 264.7 macrophage-like cells. Methods and Results : The results showed that Ganoderma lucidum inhibited inducible NO synthase (iNOS) expression of RAW 264.7 macrophage-like cells at non-cytotoxic concentrations probably through the reduction of reactive oxygen species (ROS) production. After pre-treatment of cells with non-toxic doses of Ganoderma lucidum; NO production was significantly decreased. Moreover, Ganoderma lucidum treatment suppressed LPS/IFN-γ -stimulated pro-inflammatory cytokine secretion, including interleukin-1β and interleukin-6, in a dose-dependent manner. Conclusion : Taken together, these results indicate that the anti-inflammatory activation of Ganoderma lucidum in LPS/IFN-γ-stimulated macrophages might be due to abrogation of NO-dependent cytokine release by impairment of iNOS expression via ROS generation.

This study examined the effects of trifoliate orange extract (TOE) on inflammatory reactions at the time of an LPS shock by performing experiments on rats injected with trifoliate orange extract and in Raw 264.7 cell cultures, with the aim of developing a new anti-inflammatory medicine. The IL-1β, IL-6, and TNF-α concentrations were lower in all of the groups treated with TOE than in the control group after 5 h of LPS treatment. The IL-10 concentration was higher in the 300- ㎎/㎏ TOE group than in the control group after 2 h and 5 h of LPS treatment. The liver concentrations of cytokines IL-1β and IL-6 decreased more in the groups treated with TOE than in the control group and the IL-6 concentration did not differ significantly between the 100-㎎/㎏ TOE group than in the control group. The TNF-α and IL-10 concentrations did not differ significantly between the TOE groups and the control group. In the experiments involving Raw 264.7 macrophage cultures subjected to LPS shock, the productions of IL-1β, IL-6, and TNF-α decreased in all of the groups treated with TOE compared to the control group. The IL-10 concentration did not differ significantly between the groups treated with TOE and the control group. Together the findings of this study suggest that TOE contains functional substances that can influence inflammatory reactions.

Background: Sedum takesimense Nakai has been used as folk medicine in Korea. The present study aimed to determine the biological activity of S. takesimense by investigating the anti-inflammatory effects of S. takesimense water extract (SKLC) on the lipopolysaccharide-induced inflammatory response in RAW 264.7 cells. Methods and Results: Cytotoxicity of SKLC on RAW 264.7 cells was determinded by performing MTS assay was found to have no cytotoxic effect on RAW 264.7 cells at a concentration range of 62 - 500 ㎍/㎖. Further, pretreatment of SKLC inhibited lipopolysaccharide-induced nitric oxide (NO) production in a dose-dependent manner. To determined the inhibitory mechanisms of SKLC on inflammatory mediators, we assessed the inducible nitric oxide synthase (iNOS) and cyclooxygnease-2 (COX-2) pathways. The activities of these pathways were decreased in a dose-dependent manner by SKLC. The production of tumor necrosis factor- α (TNF-α), interleukin (IL)-1β‚ and IL-6 were also reduced. Conclusions: These results suggest that the down regulation of iNOS, COX-2, TNF-α, IL-1β‚ and IL-6 expression by SKLC are mediated by the down regulation of nuclear factor-κB (NF-κB) activity, a transcription factor necessary for pro-inflammatory mediators. This might be the mechanism underlying the anti-inflammatory effects of SKLC.

This study describes a preliminary evaluation of the anti-inflammatory activity of Acrosorium yendoi Yamada extracts. A. yendoi Yamada was extracted using 80% ethanol and then fractionated sequentially with n-hexane, ethyl acetate and butanol. To screen for anti-inflammatory agents effectively, we first examined the inhibitory effect of 80% EtOH extract and solvent fractions of A. yendoi Yamada on the production of pro-inflammatory factors and cytokines stimulated with lipopolysaccharide. In addition, we examined the inhibitory effect of 80% EtOH extract and solvent fractions of A. yendoi Yamada on pro-inflammatory mediators (NO, iNOS, PGE2, and COX-2) in RAW 264.7 cells. In the sequential fractions of n-hexane and EtOAc inhibited the NO and PGE2 production and the protein level of iNOS and COX-2, and protein expression of pro-inflammatory cytokines (TNF-α, and IL-6). These results suggest that A. yendoi Yamada may have significant effects on inflammatory factors and may be provided as possible anti-inflammatory therapeutic seaweed.

Arabis glabra is a localized common rhizomatous flowering plant, This plant is often used in Korean traditional systems of medicine as a remedy for blood cleaning, detoxification, abscess, gastrospasm, arthritis, contraction and diarrhea. Generally drugs that are used for arthritis have antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of A. glabra using the ethanolic extract and its sub-fractions. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) on RAW 264.7macrophages. Our results indicated that hexane and chloroform fraction significantly inhibited the LPSinduced NO and PGE2 production in the cells. The hexane fraction inhibitory activity for NO tests with IC50 values showed in 21.0 ㎍/㎖. The chloroform fraction inhibitory activity for PGE2 tests with IC50 values showed in 18.0 ㎍/㎖. These efficacy are expected to be able to present the potential for the development of health functional food for the prevention inflammatory diseases because it has sufficient preventive medical possibilities. Further, it is determined that it is necessary to further study the mechanism of cytokine and protein expression associated with inflammation.

다양한 제품들에 사용되고 있는 천연물 소재들의 효능에 대한 체계적이고 과학적인 증거자료와 임상자료는 매우 부족한 실정이다. 이러한 천연물 소재의 과학적 연구는 국 민보건과 건강증진을 위한 다양한 제품개발의 기초자료로 활용할 수 있는 중요한 자료이다. 따라서 본 연구에서는 보리순 추출물을 이용하여 기능성 천연물 소재로서의 가능 성을 검토하였다. 항염증 활성을 조사하기 위해서 대식세 포 RAW264.7에 LPS로 자극시켜 유도된 염증반응에서 보 리순 에탄올 추출물의 매개체 억제 효과를 수행하였고 oxazolone을 이용하여 hairless 마우스에 접촉성 피부염을 유도하여 보리순 추출물의 항염 효과를 확인하였다. 연구 의 결과에서, LPS로 자극한 RAW264.7 세포에서 COX-II, iNOS와 같은 염증성 매개체뿐만 아니라 염증성 사이토카 인의 생성 및 발현이 보리순 에탄올 추출물에 의하여 현저 히 억제됨을 확인하였다. 이러한 보리순 에탄올 추출물의 억제효과는 IκB의 분해반응을 억제함으로써 NF-κB의 핵 으로 이동을 억제하여 신호전달체계를 불활성화시키는 것 과 관련이 있는 것으로 나타났다. 또한 보리순 에탄올 추출 물은 NF-κB의 상위 신호전달경로인 MAPKs에도 영향을 미치는 것으로 증명되었다. 대식세포에서의 실험결과를 토 대로 hairless 마우스에 oxazolone으로 접촉성 피부염을 유 발시킨 모델에서 보리순 에탄올 추출물을 처리하여 2주간 피부 병변을 관찰한 결과 염증반응이 현저히 감소됨을 확인 하였다. 이상의 결과에서 보리순 추출물이 항염증 효능을 가진 천연물 소재로서의 가능성을 제시하고 있다. 기존 연 구에 의하면, 보리 추출물에는 항산화 효과가 뛰어난 폴리 페놀류인 루테오린(luteolin), 사포나린(saponarin) 등이 풍 부하며, 항염증 효과가 우수한 페루릭산(ferulic acid), 루토 나린(lutonarin), 그리고 각종 비타민, 미네랄 등이 풍부하다 고 알려져 있다. 또한 superoxide와 hydroxyl 라디칼 생성을 억제할 수 있는 2"-O-glycosylisovitexin이 함유되어 있는 것으로 밝혀졌다. 이러한 성분들에 의하여 보리순 추출물 의 항염 효과가 나타날 것으로 생각된다. 다양한 염증성 질환에서 여러 매개체들의 과도한 발현이 그 질환의 원인임 을 생각해 볼 때 보리순 추출물은 항염증에 관련된 여러 제품들에서 다양하게 활용할 수 있는 천연물 소재가 될 것으로 판단되며 앞으로 보리순 추출물에서 항염 효과를 나타내는 유효화합물의 분리동정 및 생리활성평가에 대한 연구가 진행되어야 할 것으로 사료된다.

본 연구에서는 마늘의 부산물로 발생하는 마늘대의 항산화 및 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 Raw 264.7 세포에 대한 열수추출물(ASSW)과 70% 에탄올 추출물(ASSE)의 효과를 살펴보았다. ASSW의 폴리페놀 함량은 37.08±1.51 mg(TAE)/g, ASSE의 폴리페놀 함량은 44.7±1.32 mg(TAE)/g 이 함유되어있음을 확인하였다. DPPH 실험과 ABTS+ 실험에서 ASSW, ASSE 모두 농도 의존적으로 증가하였으며, DPPH의 경우 1,000 μg/mL에서 대조군인 Vit.C의 50 μg/mL의 항산화능이 있다는 것이 확인되었고, ABTS+의 경우 500 μg/mL 이상부터 대조군인 Vit.C와 비슷한 효과를 나타냄으로서 ASSW, ASSE 모두 항산화능이 있다는 것을 확인하였다. MTT측정으로 인해 세포 독성을 가지지 않았던 농도대(5, 10, 25, 50, 100 μg/mL)에서 염증 억제 효과를 살펴보기 위해 NO를 측정한 결과 ASSW, ASSE 모두 25 μg/mL에서부터 유의적으로 분비량이 감소함을 확인하였으며 특히 100 μg/mL의 농도에서 약 18%, 23%의 억제 효과를 보였다. 또한 대식세포의 염증성 cytokine인 IL-6, TNF-α, IL-1β 및 PGE2의 분비량을 첨가 농도 의존적으로 억제함을 확인하였다. 특히 PGE2에 대해 ASSW, ASSE 100 μg/mL의 농도에서 약 55%, 60%의 감소효과를 보였다. ASSW의 iNOS, COX-2의 발현 저해효과는 나타내지 못하였지만, ASSE는 100 μg/mL의 농도에서 iNOS의 발현량이 현저하게 억제됨을 확인하였고, COX-2의 경우 농도 의존적으로 저하되어 특히 50 μg/mL, 100 μg/mL의 구간에서 단백질 발현 저해효과가 있음을 확인하였다. 이를 통해 ASSW, ASSE 모두 항산화 효과와 항염증 효과가 있음을 확인하였으며, ASSW 보다 ASSE에서 활성산소종(reactive oxygen species, ROS) 및 ROS에 의해 유발되는 염증을 억제시켜주는 소재가 될수있을 것이라 예상된다.

Anti-inflammatory activity of the extracts of ginseng berry (GBE) was investigated through the evaluation of its inhibitory effect on the production of inflammatory meditator, nitric oxide(NO), tumor necrocis factor-alpha (TNF-α), interleukin-6 (IL-6) in LPS-induced RAW264.7 macrophage cells. GBE was fractionated using n-hexane, chloroform, ethylacetate, buthanol and H2O, sequentially. RAW264.7 cells were induced 100ng/mℓ of lipopolysaccharide (LPS) and treated with 0, 1.6, 8, 40 and 200μg/mℓ of GBE fractions. LPS-induced NO production on all of GBE fractions was inhibited with increasing added concentration of GBE fractions. Chloroform fraction of GBE was the most effective in inhibiting LPS-induced TNF-α production. Hexane, chloroform and H2O fractions of GBE exhibit strong inhibition LPS-induced IL-6 production. Especially, H2O fractions of GBE was the most effective in inhibiting LPD-induced IL-6 production without significant cytotoxicity in RAW264.7 cells, and reduced the activation of mitogen-activated protein kinases (MAPK) and IkB phosphorylation. These results indicate that H2O fractions of GBE exhibits strong anti-inflammatory effects by inhibition of NF-kB by inhibition of p-38 on MAPK and IkB phosphorylation.

적하수오 추출물의 추출용매별 항염증 작용을 알아보기 위해 LPS로 자극된 RAW 264.7 세포를 이용하여 실험을 진행 한 결과 열수 추출물 및 70% 에탄올 추출물 모두 100μg/mL에서 60%이상의 NO 생성 억제 율을 보이는 것을 확인 하였다. 또한 사이토 카인들에 대하여 적하수오 추출물은 TNF-α, IL-1β, IL-6, PGE2 생성량은 농도 의존적으로 감소하였고, 이는 적하수오 추출물이 염증성 사이토 카인의 발현을 억제 하여 염증 발생을 억제 할 수 있다고 할 수 있다. PMW는 100μg/mL의 농도에서 iNOS 단백질 발현량이 89%, COX-2는 54%의 효과를 나타내었으며, PME는 100μg/mL의 농도에서 iNOS 91%, COX-2는 57%의 단백질 발현 저해효과가 있음을 확인 할 결과로 보아 적하수오는 대식세포에서 NO, TNF-α, IL-1β, IL-6, PGE2 iNOS, COX-2 발현을 억제 하며, 특히 적하수오 70% 에탄올 추출물의 경우 저 농도에서부터 효과를 나타내어 우수한 항염증 소재임을 확인 할 수 있었다.