국내자생 야생버섯의 항염증 활성검증을 위해 7월~8월에 채집된 버섯을 메탄올 추출하여 DMSO의 희석한 후 Nitric oxide 저해능을 분석하였다. 30여종의 버섯 추출물을 세포활력도를 측정하기 위해 MTT 분석결과 12개의 버섯추출물이 비교적 안정된 세포활력도를 보였으며, nitric oxide 저해능을 양성대조군(LPS)와 비교한 결과 Amanita virgineoides:흰가시광대버섯, Tylopilus neofelleus:제주쓴맛그물버섯, Armillariella tabescens:뽕나무버섯부치, Amanita abrupta:양파광대버섯, Agaricus subrutilescens:진갈색주름버섯, Boletus subvelutips 7종의 버섯은 18%이상의 우수한 nitric oxide저해능을 보였다.

Atopic dermatitis is a chronic inflammatory skin disease associated with cutaneous hyper-reactivity to environmental triggers. In order to develop effective therapeutic herbal extracts for atopic dermatitis, cytotoxicity, antioxidant, anti-inflammatory and anti-allergic activities were investigated for various herbal extracts. Among candidate extracts, we selected Aloe vera L. (AV), Viola mandshurica W. Becker (VM), Punica granatum L. (PG), Dendrobium nobile L. (DN) and mixture of the above extracts (MX) for further investigations. All of them did not show cytotoxic activities to macrophage RAW264.7 cells below the concentration of 100 ppm. All showed antioxidant, anti-inflammatory and anti-allergic effects, although to various extents. In antioxidant effects, AV showed the highest effect, followed by PG and VM, while DN did the lowest. In evaluation for anti-inflammatory activities in macrophage RAW264.7 cells, AV and DN inhibited almost completely the LPS-induced production of nitric oxide, while AV, DN and VM showed strong inhibitory activities on the LPS-induced production of TNF-α. In anti-allergy effect in mast cell HMC-1, DN showed the highest effect, followed by AV and PG, while VM did the lowest. In the topical allergy reaction induced by compound 48/80 in Sprague-Dawley rat, DN exhibited significant anti-allergic effect, while PG, VM and AV did slight effect. These results suggest that AV, VM, PG and DN have antioxidant, anti-inflammatory and anti-allergic activities, and thus have the potential to reduce and alleviate the symptoms of atopic dermatitis.

This study was carried out to investigate the effects of Hericium erinaceus extract on the Anti- inflammatory and Anti-bacterial. To investigate in vitro Anti-bacterial activity assay, hot water extracts of Hericium erinaceus carried out using a broth micro-dillution method following the Clinical and Laboratory Standards Institute guidlines(Citron and Hecht, 2003; Jorgensen and Turnidge, 2003). In the next experiment, to investigate anti-inflammatory test, the RAW 264.7 macrophage, cells was cultured using DMEM including the 10% FBS. Hericium erinaceus extract has the effects of anti-inflammation and, which may be due to its inhibitory potential on the macrophage activation. Furthermore, Hericium erinaceus extract has the anti-bacterial effects through the inhibitory Propionibacterium acnes(P. acnes) and periodontopathic bacteria, Porphyromonas gingivalis , Fusobacterium nucleatum and Tannerella forsythia. The above results suggest that Hericium erinaceus hot water extract could be applicable for improvement of several inflammatory disease.

Phellinus gilvus(PG) is a medicinal mushroom belonging to the Hymenochaetaceae basidiomycetes, and has advantages over many Phellinus species due to its short growth period (3 mo), making it cheaper to produce. In the current investigation, we determined the major components of the ethyl acetate extract of PG responsible for its biological activities and further compared the magnitude of the antioxidant/anti-inflammatory activities of components with the various fractional extracts of PG. As the results, the average total DPPH radical scavenging activities of both Fd and Fc of PG was 10 mg/mL, > 95%. Among the fractional extracts of PG, Fd had the greatest inhibitory activity with an IC50 value of 36.70㎍/mL, whereas Fb showed the lowest activity. PCA had even greater activity of NO inhibition than Fd with an IC50 value of 19.46㎍/mL. The mRNA expression of iNOS or COX-2 was nearly undetectable in the absence of LPS. However, LPS- stimulation markedly increased the expression of both iNOS and COX-2 genes. Fd inhibited the effect of LPS in a concentration-dependent manner. Six major compounds were identified from the ethyl acetate extract of PG, and protocatechualdehyde (PCA) was supposed to be the major phenolic compound of PG responsible for its DPPH free radical scavenging activity and its inhibitory effects on LPS-induced NO production in RAW264.7 cells. Further in vitro and in vivo experiments are currently underway to confirm this observation and to investigate the detailed molecular mechanisms involved in the process as well as the biological activities of other fractions of Fd.

Anti-inflammatory effects of ethanolic extract of Angelica gigas (EAG 50, 150, and 500 mg/kg) were investigated in thermal burn-paw edema model. Immersion of rat hind legs into 48.5℃ water for 50 scincreased the leg volume by 0.46-0.55 ml. EAG attenuated not only the paw volume in a dose-dependent manner, but also blood lymphocyte counts and interleukin-6 level increased by burns, without influence on the level of tumor-necrosis factor-α. Moreover, EAG reduced alkaline phosphatase level, a parameter of hepatic injury, which was increased by thermal burns. The results indicate that EAG attenuates inflammatory aymptoms induced by thermal burn and EAG or its conmarin derivatives could be a promising anti-inflammatory drug candidate in inflammation diseases.

염증은 신체 특정 조직의 감염 및 손상에 관한 생체 반응이며, 매개하는 주요 대상은 면역세포이다. 염증은 급성과 만성 염증으로 나뉘며 신체 조직의 감염 및 손상부위의 규모에 따라 구분 할 수 있다. 염증의 범위가 크게 발현되거나 급성염증 형태로 진행되지 않을 때 만성 염증으로 진행되며 대표적인 만성 염증 질환인 장 질환(Inflammatory bowel disease)의 일종인 크론병 (Crohn’s disease)이나 관절질환인 류머티스성 관절염(Rheumatoid arthritis)으로 나타난다. 낮은 수준이기는 하나 비만 역시 염증성 질환으로 분류할 수 있다. 연리초속 식물이 고래에 신장염을 치료하는 민간처방으로 주로 사용됐기에 이에 착안하여 털연리초(Lathyrus palustris)를 이용하여 세포독성과 항염증 활성 효과를 평가하였다. 대식세포인 RAW 264.7 세포에서 염증 유발 인자인 lipopolysaccharide (LPS)로 자극 후 NO와 PGE2 같은 염증 매개 물질들의 억제 효과를 확인하였다. 털연리초 에탄올 추출물을 처리한 후 염증 매개 물질의 저해율(%)을 측정했을 때 NO 및 PGE2 생성을 농도 의존적으로 현저하게 억제하는 농도는 40 ㎍/mL이었으며 특이적으로 PGE2 발현을 74% 이상 강력히 억제함을 확인하였다. 따라서 본 연구결과는 털연리초의 에탄올 추출물이 유의성 있는 항염증 효과를 나타내었고 이러한 생리활성 효과는 예방의학적 소재로서의 가능성을 충분히 제시할 수 있기에 염증 질환의 예방 및 비만 억제를 위한 기능성 건강식품의 개발로 이어질 것으로 기대된다. 또한 염증 과 관련된 사이토카인 물질인 IL-4, IL-13 및 염증 지표 단백질 인 iNOS, COX-2의 억제 메커니즘과 항염증 활성을 나타내는 핵심 성분의 추가적인 연구가 차후 필요할 것으로 판단된다.

The root extracts of Paeonia lactiflora cv. ‘Red Charm’ has been studied by many groups, however, little attention has been paid to its flower petal. Paeonia is the genus in the Paeoniaceae family. ‘Red Charm’ Paeonia is a soft-stemmed herbaceous peony hybrid of P. officinalis and P. lactiflora. We previously showed the flower petal extract of Red Charm might have anti-oxidant and anti-inflammatory activities, however, it was not clear which components might be involved in this activity. Bioinformatics analysis previously indicated these extracts have potential anti-oxidant materials. One of them is identified as paeoniflorin, which is major component in root extract of Red Charm. In this study, we compared paeoniflorin and oxypaeoniflorin using DPPH assays to measure its anti-oxidant activities. Oxypaeoniflorin showed higher levels of radical scavenging activity, similar to ascorbic acid control, whereas paeoniflorin did not. Furthermore, nitric oxide assay showed they have similar anti-inflammatory effects. Taken together, these results suggest oxypaeoniflorin may play a more important role in the anti-oxidant activity of the flower petal and root extracts of Red Charm, compared to paeoniflorin. Further studies may be able to provide a platform to develop potential dual effects therapeutics for oxidant-mediated and inflammation-mediated disease in the near future.

Quercus mongolica (QM), which belongs to fagaceae, is one of the oak native to Korea. We evaluated the antiinflammatory effect of branches extracted with 70% ethanol of QM (QM-B) and elucidated the potential signaling pathway in LPS-induced RAW264.7 cells. The QM-B showed anti-inflammatory activity through inhibition of NO production. The QM-B dose-dependently suppressed NO production by inhibiting iNOS, COX-2 and IL-6 expression in LPS-induced RAW264.7 cells. The QM-B inhibited the degradation and phosphorylation of IκB-α and NF-κB activation. The QM-B suppressed the phosphorylation of p38 and ERK1/2. Also, the QM-B increased HO-1 expression. These results suggested that QM-B may utilize anti-inflammatory activity by suppressing NF-κB and MAPK signaling pathway and inducing HO-1 expression indicated that the QM-B can be used as a natural anti-inflammatory drugs.

Zanthoxylum piperitum D.C. (ZP) peels has been used as a natural spice and herb medicine for hypertension reduction, for strokes, and for its anti-bacterial and anti-oxidant activity. However, the anti-inflammatory mechanisms employed by ZP have yet to be completely understood. In this study, we elucidate the anti-inflammatory mechanism of ZP in lipopolysaccharide (LPS)-induced RAW264.7 cells. We evaluated the effects of ZP in LPS-induced levels of inflammatory cytokines, prostaglandin E2 (PGE2), and caspase-1 using ELISA. The expression levels of inflammatoryrelated genes, including cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS), were assayed by Western blot analysis. We elucidated the effect of ZP on nuclear factor (NF)-κB activation by means of a luciferase activity assay. The findings of this study demonstrated that ZP inhibited the production of inflammatory cytokine and PGE2 and inhibited the increased levels of COX-2 and iNOS caused by LPS. Additionally, we showed that the anti-inflammatory effect of ZP arises by suppressing the activation of NF-κB and caspase-1 in LPS- induced RAW264.7 cells. These results provide novel insights into the pharmacological actions of ZP as a potential candidate for development of new drugs to treat inflammatory diseases.

Coriandrum sativum L., an annual herbaceous plant of Apiaceae family. The present study evaluated the anti-oxidant activities and anti-inflammatory effects of ethanol extracts of C. sativum. The anti-oxidant activities of C. sativum were measured by total contents of polyphenol, flavonoid, DPPH and ABTS radical scavenging and reducing power activity. And anti-inflammatory effects of C. sativum were measured by LPS-induced RAW 264.7 cells. The results showed that the contents of total polyphenol and flavonoid were 76.03 ± 1.36 mg of gallic acid equivalents/g and 182.23 ± 4.32 mg of rutin equivalents/g at concentration 1 mg/ mL of C. sativum. The DPPH radical scavenging activity was found to be 52.8% at 500 μg/mL. The ABTS radical scavenging activity was shown in 58.3% after exposure to 1,000 μg/mL. Reducing power activity was found to be 66.8% at 2,000 μg/mL. The inhibitory effect of NO production was found to be 65% concentration 500 μg/mL. In the generation quantity of inflammatory cytokines such as TNF-α and IL-1β in cell culture medium, the expression levels of inflammatory proteins in cells were showed decrease with the increase of concentration. Therefore, we suggest that the C. sativum should be a potential source of alternative anti-inflammatory drug with good anti-inflammatory effects.

본 연구는 하라케케(Harakeke)로 불리는 신서란(Phormium tenax)를 화장품 및 의약품산업의 기능성 소재로서의 이용 가능 성을 확인하기 위하여 신서란 잎을 대상으로 70% 에탄올 추출물과 용매 분획물을 제조하여, 이것들의 항염증 및 항아토피의 효과를 조사하였다. LPS로 유도된 RAW 264.7 세포에서 신서란 에탄올 추출물과 용매 분획물의 항염증 효과를 조사한 결과, methylene chloride와 ethyl acetate 분획물에서 NO와 PGE2 생성 억제 활성이 가장 높게 나타났으며, 농도 의존적으로 NO와 PGE2 생성 억제 활성을 보였다. 또한, 이들 분획물에서는 iNOS 및 COX-2 발현 억제 활성을 보였다. 신서란 잎 조추출물과 용매 분획물에 의한 NO, PGE2 생성 억제 활성이 NOS 및 COX-2 발현 억제에 의한 것임을 제시한다. 더불어, hIFN-γ로 자극된 HaCaT 세포에 용매 분획물을 처리하여 MDC 및 TRAC 생성억제 효과를 조사한 바, methylene chloride 분획물은 MDC 및 TATC의 생성을 각각 65%, 52% 생성억제 시켰으며, ethyl acetate 분획물은 MDC 및 TATC의 생성을 각각 93%, 84% 억제 효과를 보였다. 이상의 결과는 신서란 잎 조추출물과 용매 분획물을 이용한 항염증 및 항아토피 효능을 갖는 유효성분 분리 및 활용화 연구에 중요한 기초자료가 될 것이며, 기능성 화장품, 의약외품 및 의약품 소재 개발에 적용 가능성이 높다고 사료 된다.

쉽싸리(L. lucidus)가 가지는 효능의 하나로 알려진 항염증 효과의 활성물질을 파악하기 위하여 본 연구를 수행하였다. 항염효과는 LPS로 활성화한 macrophage 264.7이 생산하는 NO의 감소효과를 측정함으로써 평가하였다. 쉽싸리 추출물에서 얻은 비극성 분획물인 CHCl3 분획물은 농도의존적으로 현저히 NO 생산을 감소시켰다. 이에 비해 극성 분획물인 BuOH 분획물은 그 효과가 약하였다. Silica gel column chromatography에 의해 이 CHCl3 분획물로부터 주요 화합물인 ursolic acid를 분리하고 분광학적 방법으로 동정할 수 있었다. 효과가 약하였던 BuOH 분획물로부터 diaion HP-20 column chromatography와 sephadex LH-20 column chromatography로 이 분획의 주요 화합물인 rosmarinic acid를 분리하고 역시 분광학적 방법으로 동정하였다. Ursolic acid는 농도의존적으로 NO 생산을 억제하였으나 rosmarinic acid는 그 효과가 상대적으로 약하였다. 이러한 사실로부터 쉽싸리의 항염효과는 주로 ursolic acid의 존재 때문임을 알 수 있었다.

최근 미세먼지, 내분비장애, 환경 호르몬등에 의해 피부 염증 질환 환자들이 증가함에 따라 화장품 소재 개발 연구는 스테로이드, 항히스타민제 등의 피부염 치료 합성화합물 보다 이를 대체할 수 있는 천연물 기반의 보다 안전한 소재 발굴에 관심이 더 증가되고 있다. 따라서 본 연구에서는 여러 가지 천연 약재 스크리닝을 거쳐 피부 효능 효과가 검증된 두충, 익지인, 백강잠의 한방추출물(Mix)을 선정하고, 이들의 새로운 피부 효능을 규명하여 천연 화장품 소재를 개발하고자 하였다. 우선 한방추출물(Mix)의 항산화 효능을 검증하기 위해 DPPH assay를 수행한 결과 400 μg/mL 농도 조건에서 약 98.8% DPPH radical 소거 활성을 확인하였다. 한방추출물(Mix)의 미백효능을 확인하기 위해 B16F10 세포주를 활용하여 한방추출물에 의한 멜라닌 합성량을 측정하였다. 400 μg/mL 농도 조건에서 α-MSH 처리군 대비 27.1% 수준까지 멜라닌 합성량이 감소하여 미백 효능 활성에 대해 유의성 있는 결과를 확인하였다. 또한 한방추출물(Mix)의 항염증 효능을 확인하기 위해 LPS 자극으로 활성화된 RAW264.7 대식세포에서 NO(nitric oxide) 합성 및 iNOS, COX-2의 유전자 발현을 관찰한 결과 모두 유의적으로 감소되는 것을 확인하였다. 염증성 사이토카인으로 알려진 IL-1α, IL-1β, IL-6, TNF-α의 mRNA 발현을 분석한 결과에서도 한방추출물에 의해 효과적으로 억제되는 것을 확인하였다. 마지막으로 피부장벽 기능 강화를 확인하기 위해 인간 각질형성세포인 HaCaT을 이용해 tight junction 구성 단백질 인 claudin 1 유전자의 발현 변화를 관찰한 결과 한방추출물에 의해 mRNA 발현이 농도 의존적으로 증가되는 것을 확인하였다. 따라서 본 연구를 통해 우리는 두충, 익지인, 백강잠을 함유한 한방추출물(Mix)이 항산화, 항염, 미백, 그리고 피부장벽 강화 등에 유효한 소재임을 확인하였고, 향후 피부 임상시험 등이 이뤄진다면 피부염 환자에 효과적인 천연 화장품 소재로 개발될 수 있을 것이다.

Aralia cordata (A. cordata), which belongs to Araliaceae, is a perennial herb widely distributed in East Asia. We evaluated the anti-inflammatory effect of stems (AC-S), roots (AC-R) and leaves (AC-L) extracted with 100% methanol of A. cordata and elucidated the potential signaling pathway in LPS-stimulated RAW264.7 cells. The AC-L showed a strong anti-inflammatory activity through inhibition of NO production. AC-L dose-dependently inhibited NO production by suppressing iNOS, COX-2 and IL-β expression in LPS-stimulated RAW264.7 cells. AC-L inhibited the degradation and phosphorylation of IκB-α, which donated to the inhibition of p65 nuclear accumulation and NF-κB activation. Furthermore, AC-L suppressed the phosphorylation of ERK1/2 and p38. These results suggested that AC-L may utilize anti-inflammatory activity by blocking NF-κB and MAPK signaling pathway and indicated that the AC-L can be used as a natural anti-inflammatory drugs.

Honey used as conventional medicine has various pharmacological properties. In the honey and anti-inflammatory effect, Gelam honey and Manuka honey has been reported to exert anti-inflammatory activity. However, the anti-inflammatory effect and potential mechanisms of acacia honey (AH) are not well understood. In this study, we investigated anti-inflammatory activity and mechanism of action of AH in LPS-stimulated RAW264.7 cells. AH attenuated NO production through inhibition of iNOS expression in LPS-stimulated RAW264.7 cells. AH also decreased the expressions of IL-1β, IL-6 and TNF-α as pro-inflammatory cytokines, and MCP-1 expression as a pro-inflammatory chemokine. In the elucidation of the molecular mechanisms, AH decreased LPS-mediated IκB-α degradation and subsequent nuclear accumulation of p65, which resulted in the inhibition of NF-κB activation in RAW264.7 cells. AH dose-dependently suppressed LPS-mediated phosphorylation of ERK1/2 and p38 in RAW264.7 cells. In addition, AH significantly inhibited ATF2 phosphorylation and nuclear accumulation of ATF2 in LPS-stimulated RAW264.7 cells. These results suggest that AH has an anti-inflammatory effect, inhibiting the production of pro-inflammatory mediators such as NO, iNOS, TNF-α, IL-6, IL-1β and MCP-1 via interruption of the NF-κB and MAPK/ATF2 signaling pathways.