The aim of this study was to evaluate the probiotic properties of lactic acid bacteria (LAB) isolated from kimchi and antioxidant and anti-inflammatory activities of heat-killed LAB strains. Regarding probiotic properties, Lactiplantibacillus plantarum SMF398 and L. plantarum SMF470 isolated from kimchi can survive under artificial gastric condition and adhere strongly to HT-29 cells. The SMF398 and SMF470 strains showed strong antimicrobial activity and co-aggregation against pathogenic bacteria. The heat-killed cells of LAB (1 mg/mL) were prepared by heating at 121℃ for 15 min. MTT assay revealed that the heat-killed cells (1 mg/mL) of SMF398 and SMF470 were not toxic to HT-29 cells. The heat-killed SMF398 and SMF470 showed significantly higher DPPH and ABTS radical scavenging activities as well as β- carotene bleaching inhibitory activity than the heat-killed L. plantarum ATCC14917, a control probiotic strain (p<0.05). In lipopolysaccharide-induced RAW264.7 cells, the heat-killed SMF398 and SMF470 significantly reduced the nitric oxide production by 30.92% and 22.81%, respectively (p<0.05). Furthermore, the heat-killed SMF398 and SMF470 significantly decreased the gene expression levels of tumor necrosis factor-α, interleukin-6, inducible nitric oxide synthase, cyclooxygenase-2 up to 58.02%, 67.70%, 53.44%, 45.90%, respectively (p<0.05). These results suggest that the heat-killed L. plantarum SMF398 and L. plantarum SMF470 might be useful as antioxidant and anti-inflammatory agents.
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.
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.
본 연구에서는 당근 지상부 추출물 및 용매 분획물의 항염, 항균 활성을 확인하고 유효성분을 분리하여 화학구조를 동정하였다. 당근 지상부 에탄올 추출물 및 용매 분획물의 항염 활성을 측정하기 위해 LPS로 자극된 RAW 264.7 세포를 이용하여 NO 생성 억제 활성을 확인하였다. 그 중 에틸아세테이트 분획물이 NO의 생성을 농도 의존적으로 감소시키는 효과가 나타났고, 추가적인 항염 기전 연구를 위해 에틸아세테이트 분획물에 대해 PGE2, 전염증성 사이토카인의 생성량 및 iNOS, COX-2 단백질의 발현량을 측정하였다. 그 결과, 에틸아세테이트 분획물이 PGE2, IL-1β, IL-6의 생성을 감소시키고, iNOS, COX-2 단백질의 발현도 억제 시키는 효과가 있음을 확인하였다. 또한 표피포도상구균과 여드름균을 이용한 활성 실험 결과, 헥산 및 에틸아세테이트 분획물에서 항균 활성이 나타났다. 이와 같은 결과를 바탕으로 에틸아세테이트 분획물에 대해 컬럼 크로마토그래피를 수행하여 3개의 화합물을 분리하였으며, 1H 및 13C NMR 데이터 분석과 문헌을 통하여 화학구조를 동정하였다; diosmetin (1), disomin (2), cynaroside (3). 분리된 화합물 1-3에 대해 항염 및 항균 활성을 측정하였으며, 그 결과 화합물 1-3 모두 NO의 생성을 저해시키는 효과가 있음을 확인하였다. 또한 화합물 3은 여드름균에 대해 항균 활성이 우수하였다. 이상의 연구 결과를 바탕으로 당근 지상부 추출물을 이용한 항염 및 항균 효과를 갖는 천연 화장품 소재로의 개발이 가능할 것이라 사료된다.
Mushrooms have been widely cultivated and consumed as foods and herbal medicines owing to their various biological properties. However, few studies have evaluated the anti-inflammatory effects of mushrooms. Here, we investigated the effects of mushroom extracts (MEs) on lipopolysaccharide (LPS)-induced inflammation in macrophages (RAW264.7 cells). First, we extracted MEs with either water or ethanol. Using LPS-treated RAW264.7 cells, we measured cell proliferation and NO production. Gene expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 (IL-6), and IL-1β was assessed by RT-PCR, and protein abundance of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) and phosphorylation of p65 were determined by immunoblotting. MEs prepared using both water and ethanol inhibited LPS-induced inflammation in RAW264.7 cells. Nitric oxide (NO) levels induced by LPS were reduced by treatment with MEs. Isaria japonica Yasuda water extracts and Umbilicaria esculenta (Miyoshi) Minks ethanol extracts significantly decreased the mRNA expression of inflammation-related cytokine genes including TNF-α, IL-6, and IL-1β. Similarly, the protein abundance of iNOS and COX-2 was also decreased. The phosphorylation of p65, a subunit of nuclear factor-κB was at least partly suppressed by MEs. This study suggests that mushrooms could be included in the diet to prevent and treat macrophage-related chronic immune diseases.
In this study, the anti-inflammatory activities of the extracts of different parts of Hovenia dulcis such as leaves, stems, and roots were investigated. Among them, the roots extract (RE) showed the most potent suppressive effect against pro-inflammatory mediators in LPS-stimulated mouse macrophage cells. RE induced dose-dependent reduction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and concomitantly reduced the production of NO and PGE2. Additionally, pre-treatment with RE significantly suppressed the production of inflammatory cytokines, such as tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6, as well as mRNA levels. Moreover, phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear translocation of nuclear factor-kappa B (NF-kB) were also strongly attenuated by RE in RAW264.7 cell. Furthermore, RE induced HO-1 expression through nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) and increase HO-1 activity in RAW264.7 macrophages. Therefore, these results indicate that RE strongly inhibits LPS-induced inflammatory responses by blocking NF-kB activation, inhibiting MAPKs phosphorylation, and enhancing HO-1 expression in macrophages, suggesting that RE of H. dulicis and a major component, 27-O-protocatechuoylbetulinic acid could be applied as a valuable natural anti-inflammatory material.
Background: Atractylodes radix is a well-known medicinal crop having many physiological effects. This study was conducted to select useful Atractylodes japonica × Atractylodes macrocephala (AJM) cultivars by comparing anti-oxidative and anti-inflammatory efficacies.
Methods and Results: Seven extracts from AJM cultivars were used to treat lipopolysacchride (LPS)-treated BV2 cells, and the effects on cell viability and inhibition on reactive oxygen species (ROS) and nitric oxide (NO) production were analyzed. In vitro scavenging activities of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and peroxynitrite (NOO−) radicals were also investigated. Contents of total phenol, atractylenolide I, and atractylenolide III in the AJM extracts were measured using high performance liquid chromatography (HPLC) or spectrophotometry. The experiments show that none of the seven extracts was cytotoxic above 89.2% at 20 - 250㎍/㎖. Extracts of Gowon, Dawon, Sangchul, and Huchul inhibited ROS generation in a dose-dependent manner, and Sangchul extract showed the highest inhibition on ROS production. All the AJM extracts showed effective inhibitory activity after on NO release in the LPS-treated BV2 cells, and Sangchul extract showed the highest activity. Sangchul extract had the most potent scavenging activities for NOO− and had some DPPH radical scavenging effect. Sangchul extract also had the highest content at total phenol and atractylenolide I content. Atractylenolide III was not detected in the AJM extracts.
Conclusions: The results suggested that Sangchul was the most useful anti-oxidative and anti-inflammatory resource among the AJM cultivars.
Background : It is known that Platycodon grandiflorum has anti-inflammatory activity and inhibits the production of nitric oxide (NO) in inflammatory macrophages. But the change of bioactivity of platycodon grandiflorum according to steaming is not well known. In this study, We investigated the effects of steaming on anti-inflammatory activity of 70% ethanol extracts of platycodon grandiflorum.
Methods and Result : The cytotoxicity of RAW264.7 cells treated with platycodon grandiflorum extracts was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the concentration of NO in culture supernatants were determined using nitric oxide (NO) assay. And western blotting was performed to quantify the expression of iNOS, a protein related to NO production. As a results, it was confirmed that no cytotoxicity was observed at 25, 50 and 100 ㎍/㎖ platycodon grandiflorum extracts in RAW264.7 cells. The production of NO and the expression of iNOS were induced by LPS and suppressed by all platycodon grandiflorum extracts in proportion to the number of steaming in RAW264.7 cells.
Conclusion : These results suggest that a steaming process can increase anti-inflammatory activity of platycodon grandiflorum extracts.
Backgound : Inflammatory bowel diseases (IBDs) are chronic disorders that are characterized by intestinal epithelial inflammation and injury. Currently, the most employed therapies are antibiotics and anti-inflammatory drugs; however, the side effects limit long-term effectiveness.
Methods and Results : We evaluated the impact of glucose-lysine Maillard reaction products (Glc-Lys MRPs) on colitis, induced in rats by an administration of 5% dextran sulfate sodium (DSS) in drinking water. Glc-Lys MRPs ameliorate DSS-induced colitis, as determined by a decrease in disease. index activity, colon weight/length ratio, nitric oxide levels in serum, recovery of body weight loss, colon length and serum lysozyme levels. Furthermore, Glc-Lys MRPs increase the glutathione content and the activity of glutathione peroxidase, superoxide dismutase and catalase, and inhibit lipid peroxidation and myeloperoxidase activity in colon tissues. In particular, Glc-Lys MRPs suppress the mRNA level of the inflammatory cytokines and nuclear factor-κB in colon tissues.
Conclusion : This study suggests the potential of Glc-Lys MRPs in preventing or treating IBDs.
손바닥 선인장인 백년초와 천년초 열매의 75% 에탄올 추출물을 대상으로 무기물, 총 페놀 화합물 및 플라보노이드 함량을 조사하고, 항산화, 항염증 효과를 비교함으로써 손바닥 선인장에 대한 과학적 자료를 확보하고 기능성 원료소재로의 이용 가능성을 타진하고자 진행되었다. 총 무기물 함량은 백년초 열매 추출물이 천년초 열매 추출물에 비해 약 2배 정도 높았으며, 무기물 중에는 칼륨의 함량이 가장 높았다. 총 페놀 화합물과 플라보노이드 함량 모두 백년초 열매 추출물에서 더 높았고 항산화 활성 평가를 위한 DPPH와 ABTS 라디칼 소거능 역시 백년초 추출물이 천년초 추출물보다 더 우수하였다. 세포 독성을 확인하기 위해 대식세포에서 MTT assay를 수행한 결과, 800 μg/mL의 이하 농도까지는 세포의 생존율에 영향을 미치지 않는 것을 확인하였다. LPS로 염증반응을 유도시킨 세포에서 2종 추출물 모두 농도 의존적으로 NO 생성을 억제하였으나, ROS 생성 억제 효과는 백년초 열매 추출물에서만 유의성 있는 결과가 도출되었다. 또한 세포내에서 염증 유발과 직접적으로 관련이 있는 염증성 cytokine 3종(TNF-α, IL-1, IL-6)의 발현을 확인한 결과 백년초 열매 추출물은 LPS 처리에 의해 증가된 cytokine 중 TNF-α를 제외한 IL-1과 IL-6의 발현량을 유의성 있게 감소시켰다. 이상의 실험 결과들을 종합해 볼 때 천년초 열매 추출물에 비해 백년초 열매 추출물이 더 높은 항산화 및 항염증 활성을 가지는 것으로 확인되었으며, 추후 유효성분 추출을 통한 항염증 물질의 작용기전 연구와 염증 억제 성분의 분리 연구에 기초 자료가 될 것으로 사료된다.
본 연구는 LPS로 유도된 RAW 264.7 세포에서 다양한 약용식품으로 사용되고 있는 올리브 잎과 가지 추출물의 항염증 효과를 확인하였다. 올리브 잎과 가지 추출물은 각각 RAW 264.7 세포에 대하여 세포독성을 나타내지 않았고, LPS 자극에 의한 NO 및 PGE2 생성을 농도 의존적으로 억제했다. 또한, 올리브 추출물은 LPS 자극으로 분비된 TNF-α, IL-1β 및 IL-6의 전염증성 cytokine의 분비량을 억제하였으며, 특히 200 μg/mL 농도에서 올리브 가지 추출물이 잎 추출물 보다 IL-6를 억제하는 것으로 나타났다. 대표적인 염증 관련 신호 전달 경로 인자인 iNOS 및 COX-2의 발현을 검토한 결과 올리브 추출물은 iNOS의 발현을 농도의존적으로 현저히 감소시키는 것으로 관찰되었으나, 각각의 올리브 추출물이 COX-2 발현에는 영향을 미치지 않은 것으로 관찰되었다. 이와 같은 결과는 올리브 각 부위별 추출물은 모두 iNOS 및 NO 조절 경로를 조절하는 것으로 사료되나 iNOS 및 COX-2 단백질 발현은 병립적이지 않을 수 있음을 제시하고 있다. 본 연구 결과로 올리브 추출물이 독성과 부작용이 적은 항염증 효능을 가진 기능성 화장품 소재로써 개발 가능성이 있다고 사료된다.
In this study, a preliminary evaluation of the antioxidant and anti-inflammatory activity of the Ficus erecta var. sieboldii (Miq.) King (FES) leaf extract has been performed to assess its potential as a natural resource for food and medicinal materials. FES was extracted using 70% EtOH and then fractionated sequentially using n-hexane, CH2Cl2, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, the inhibitory effect of the FES extracts on the production of oxidant stresses (DPPH, xanthine oxidase, and superoxide) and pro-inflammatory factors (NO, iNOS, COX-2, PGE2, IL-6, and IL-β) in the murine macrophage cell line RAW 264.7 activated with lipopolysaccharide (LPS) was examined. Among the sequential solvent fractions of FES, the CH2Cl2 and EtOAc fractions showed decreased production of oxidant stresses (DPPH, xanthine oxidase and superoxide), and the hexane and CH2Cl2 fractions of FES inhibited the production of pro-inflammatory factors (NO, iNOS, COX-2, and PGE2). The CH2Cl2 fraction also inhibited the production of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β). These results suggest that FES has a significant effects on the production of oxidant stresses and pro-inflammatory factors and may be used a natural resource for antioxidant and anti-inflammatory agents.
This study was conducted to compare anti-inflammatory effect of Robinia pseudoacacia L. using different extraction methods (water extraction, ethanol extraction and high temperature extraction). We investigated anti-inflammatory effect of Robinia pseudoacacia L. extract (RP1, water extract; RP2, ethanol extract; RP3, high temperature extract) on lipopolysaccharide (LPS)-stimulated inflammation using Raw 264.7 cell. Cells were treated with various concentrations (12.5, 25, 50, 100 or 200 ㎍/㎖) of water extract, ethanol extract and high temperature extract. Cytotoxicity was not observed on Raw 264.7 cells, LPS-stimulated production of NO (nitric oxide), PGE2 (prostaglandin E2) and cytokines (TNF-α, IL-6 and IL-1β) was reduced by RP3 treatment more than RP1 and RP2. In conclusion, these results indicated that inflammation on Raw 264.7 cells was improved by RP3. Treatment of RP3 could be used to natural medicine for improving inflammatory response. However, further experiment is required to observe how the high temperature extraction at 500℃ for 48 h influences on alteration of active ingredient in Robinia pseudoacacia L., and conducts the inflammation signal pathway on Raw 264.7 cells.