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.

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.

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.

In the present study, we investigated the anti-inflammatory effects by Alopecurus aequalis Sobol on thelipopolysaccharide (LPS)-induced nitric oxide (NO) production by RAW 264.7 cell line. Consistent with these observations,DS reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) atthe protein levels in a concentration-dependent manner. In addition, the release of tumor necrosis factor-α (TNF-α) andinterleukin-6 (IL-6) were also reduced by DS. Moreover, LPS increased expression phosphorylation of IκBα, but DS showedinhibitory effect by reducing LPS-inducible p-IκBα expression level. These results suggest that the down regulation of iNOS,COX-2, TNF-α, and IL-6 expression by DS are achieved by the downregulation of NF-κB activity, a transcription factornecessary for pro-inflammatory mediators, and that is also responsible for its anti-inflammatory effects.

Eucommia ulmoides OLIVER (EU) is a traditional Korean herbal used for the treatment of rheumatoid arthritis (RA). In the present study, the molecular pharmacology basis of its anti-inflammatory effect is revealed in this work, EU was studied in lipopolysaccharide (LPS)-activated macrophage cells (RAW 264.7) as an established inflammation model. After activation, nitric oxide (NO) production and iNOS mRNA were measured by using a colorimetric assay (Griess reagent), and reverse transcription polymerase chain reaction (RT-PCR), respectively. The change in the content of PGE2, TNFα, and IL-6 was concurrently monitored by ELISA. In results, we found that in the concentration range without showing cytotoxicity, EU produced a remarkable anti-inflammatory effect and showed a dose-dependent inhibition of LPSinduced NO production. Compared with indomethacin, EU has more potency and a specific action of NO inhibition, PGE2, IL-6, and TNF-α inhibition. These results suggest that EU may be a suitable herbal medicine to yield the greatest anti-inflammatory activity for food additives and medicine.

This work aimed to elucidate the anti-inflammatory effects of ethyl acetate fraction from Cnidium officinale Makino with a cellular system of LPS-stimulated RAW 264.7 and THP-1 cells. Some key pro-inflammatory cytokines and mediators including NO, iNOS, PGE2, COX-2, TNF-α, NF-kB p50 and NF-kB p65 were studied by sandwich ELISA and western blot analysis. Ethyl acetate fraction could significantly inhibit the production of NO, PGE2, TNF-α, iNOS and COX-2 in LPS-stimulated cell than that of single LPS-stimulated. And ethyl acetate fraction suppresses the activation of NF-kB p50 and NF-kB p65. All the results showed that ethyl acetate fraction had a good anti-inflammatory effect on LPS-stimulated RAW264.7 and THP-1 cells. Taken together, the anti-inflammatory actions of ethyl acetate fraction from Cnidium officinale Makino might be due to the down-regulation of NO, PGE2, TNF-α, iNOS and COX-2 via the suppression of NF-kB activation.

씀바귀추출물이 LPS shock 염증반응에서 항염증효과에 미치는 영향을 검토하기 위하여 생체 및 세포배양실험에서 전염증성 cytokine들의 생성과 혈액 내 생물학적 수치를 조사했다. 그 결과 LPS 염증유도 rat에서 씀바귀 추출물은 혈액 내 전염증성 cytokines 들, 즉 IL-1β, IL-6 및 TNF-α의 생산을 저해하고, 한편으로는 IL-10의 생산을 촉진하였다. 간장 IL-1β 및 IL-6의 농도는 씀바귀추출물 처리군이 대조군보다 유의하게 낮은 값을 나타내었으나, TNF-α 및 IL-10의 농도는 모든 처리군 간에 유의한 차이를 나타내지 않았다. Raw 264.7 cell의 세포배양실험에서는 TNF-α의 농도가 유의한 차이를 나타내지는 않았지만, 3개 cytokine 모두가 씀바귀 투여량이 증가함에 따라 감소하는 경향을 보였다. IL-10의 농도는 씀바귀 처리군 들 모두가 높은 경향을 나타내었다. 혈액 내 total protein 및 albumin의 농도는 대조군과 유의한 차이를 나타내지는 않았으나, 씀바귀추출물 처리 군에서 높은 경향을 보였다. 이상의 결과들을 종합해 보면 씀바귀 추출물에는 항염증반응에 관여하는 기능성물질이 내재하고 있음을 시사해 준다.

본 연구는 길경 사포닌 추출물(PGS)를 이용하여 대식세포의 면역조절능력을 평가하였으며, 탐식작용, 항암작용, 항염증 작용에 모두 유의적인 효과를 나타내었다. 특히, 본 연구에서는 농도의존적으로 매우 유의적이게 나타난 PGS의 항암작용 기전을 측정하기 위하여 암세포 독성 물질로 알려진 NO 분비량을 측정하였으며, PGS에 의해 NO의 생성을 증가함을 확인하였다. 또한, PGS가 NO 생성 억제제 NIL을 함께 처리하였을 때 항암효과가 나타나지 않게 됨을 재확인함으로써, PGS 10 μg/mL에서 나타낸 대식세포의 항암효과는 일부 NO 생성 및 분비에 의한 작용 기전임을 보여주었다. PGS의 면역조절작용 중 항염증효과 실험에서는 PGS가 염증환경에서 과도하게 분비된 NO를 다소 억제하는 경향을 보였으나, 항염증조절에서 대표적인 물질로 알려진 TNF-α 조절에는 효과를 나타내지 않았다. PGS가 염증환경에서의 TNF-α억제조절에는 영향을 미치지 않았으나 TNF-α는 항암물질로도 알려져 있으므로 향후 PGS의 항암효과에 대한 연구에서 TNF-α의 생성에 관한 연구는 NO를 매개하는 항암 효과 외에 다른 기전을 설명해줄 수 있을 것으로 보인다. 이러한 결과들은 PGS가 항암요법의 보조제 및 면역보조제로써의 활용에 개발 가능성이 있다는 것을 보여준다.