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.

LPS로 염증이 유도된 Raw264.7 macrophage에서 대추잎 분획물(Zizyphus jujuba leaf fractions; ZLFs)의 항염증 효과를 살펴 보기위해 세포독성이 나타나지 않은 1, 10, 100 μg/mL 농도 범위에서 염증매개물질인 NO, PGE2, 염증성 cytokine(TNF-α, IL-1β 및 IL-6) 생성 및 COX-2 단백질의 발현을 측정하였다. 그 결과, ZLFs(ZLWF, ZLEF, ZLBF)는 처리 농도 범위에서 효과적으로 NO, PGE2, 염증성 cytokine 생성 및 COX-2 단백질 발현을 억제하였다. 분획 용매에 따른 효과를 살펴보면 ZLWF< ZLBF< ZLEF의 순으로 높은 효과를 나타냈고, 특히 에틸 아세테이트 분획물 ZLEF은 100 μg/mL 처리 농도에서 NO, PGE2, 염증성 cytokine 생성 및 COX-2 단백질 발현 억제 효과가 LPS를 처리하지 않은 음성 대조군보다 우수하거나 비슷하여 본 연구에서 조사된 대추 잎 분획물 중 가장 뛰어난 염증 억제제 후보물질로 확인되었다.

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.

in vivo 및 in vitro에서 마황의 항염증효과를 검토하기 위하여 마황추출물을 급여한 흰쥐에게 LPS shock로 급성기 염증반응을 유발시킨 후, 혈액 및 간장의 전염증성 cytokines들의 농도를 경시적으로 조사하였으며, 한편으로는 Raw 264.7 cell에 LPS shock를 가한 후, 마황추출물이 각종 전염증성 cytokines들의 생산량에 미치는 영향을 조사했다. in vivo 실험에서, 각 처리군 별 plasma IL-1β, IL-6, TNF-α 및 IL-10 농도의 경시적 변동은 전 처리군 모두가 LPS 처리 후, 2h째 급격하게 증가하여, 5h째에 최고치를 나타내었다. Plasma IL-1β, IL-6 및 TNF-α의 농도는 LPS처리 후 5h째에서 마황 첨가군 모두가 대조군보다 낮은 값을 나타내었다. Plasma IL-10의 농도는 LPS처리 후, 2h째 및 5h째 모두 에서 마황추출물 첨가군 들이 대조군보다 높은 값을 나타내었다. Raw 264.7 macrophages를 이용한 in vitro 실험에서, IL-1β, IL-6 및 TNF-α의 농도들은 대조군보다 마황처리 군들 모두가 낮은 값을 나타내었다. IL-10의 농도는 대조군보다 마황처리군들이 다소 높은 경향을 보였으나, 유의한 차이는 아니었다. 이상의 결과들을 종합해보면 마황에 내재하는 기능성 물질들이 염증반응을 완화하는 효과를 가지고 있음을 시사해 준다.

A variety of herbs and plants have been traditionally used in oriental folk medicine for the treatment of inflammatory diseases. In our attempt to search for anti-inflammatory agents from natural products, we investigated 64 methanol extracts from 42 medicinal plants belonging to 10 families which were evaluated for inhibitory activities of NO production in lipopolysaccharide (LPS)-stimulated macrophage RAW 264.7 cells. Among them, 16 extracts exhibited inhibitory activities of NO production (IC50 values ranging from 59.6 to 94.7 μg/ml). Only the extract from aerial parts of Hosta lancifolia (H. lancifolia) did not exert cytotoxic effects at the concentrations tested. The extract from H. lancifolia decreased the mRNA and protein levels of inducible nitric oxide synthase (iNOS) and pro-inflammatory cytokines in activated macrophage RAW 264.7 cells in dose-dependent manner. The results suggest that the extract may contain bioactive compounds that suppress expression of pro-inflammatory cytokines, which may prove beneficial with regard to the development of natural agents for prevention and treatment of inflammatory diseases.

Rhubarb is the well-known and frequently used herbal medicine for the treatment of constipation, inflammation, and cancer. As described in the Korea and Chinese Pharmacopoeia, rhubarb consists of the roots and rhizomes of Rheum palmatum, R. officinale, and R. tanguticum. However, the pharmacological differences among rhubarb have not been scientifically established. In the present study, we investigated and compared the inhibitory effects of 70% ethanolic extracts from R. palmatum (RPE), R. officinale (ROE), R. nobile (RNE), and R. franzenbachii (RFE) on the production of inflammatory mediators, nitric oxide (NO), prostaglandin E2 (PGE2), interlukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) in RAW 264.7 macrophage cells. ROE, RNE, and RFE significantly inhibited the release of NO, PGE2, IL-1β, and TNF-α. RPE significantly reduced the release of IL-1β, but not NO, PGE2, and TNF-α. Overall, RFE was found to inhibit the release of PGE2 and IL-1β, to a far greater degree than RPE, ROE, and RNE. Our results indicate that RFE possess the strongest anti-inflammatory activity among 4 tested rhubarb.

The present study describes the preliminary evaluation of the anti-inflammatory activities of Phellinus linteus (PL) and Phellinus linteus Grow in Germinated Brown Rice (BRPL). In order to effectively screen for anti-inflammatory agents, we first examined the extracts' inhibitory effects on the expression of pro-inflammatory cytokines activated with lipopolysaccharide. Moreover, we examined the inhibitory effects of the PL and BRPL extracts on pro-inflammatory factors such as NO, iNOS, TNF-α and IFN-γ in murine macrophage RAW 264.7 cells. NO production and iNOS expression was significantly augmented in LPS treated cell, the production of NO and iNOS was greater in the BRPL than in the PL group. In addition, protein and mRNA levels of TNF-α and IFN-γ in BRPL showed relatively more potent pro-inflammatory-activity inhibition compared to that of PL. These results suggest that BRPL may have significant effects on inflammatory factors, and may be a potential anti-inflammatory therapeutic materials.

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects on the water extracts of Artemisia princeps Pampanini (APP) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased PGE2 and TNF-α production. Consistent with these results, the protein and mRNA expression level of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by water extracts of APP in a dose-dependent manner. These results suggest that APP may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

한약재 및 식용으로 사용되는 순비기나무, 번행초, 갯방풍의 재배 한계지역과 생리활성 성분의 효능은 다음과 같다. 1. 순비기나무, 갯방풍의 서해안 북방한계지역은 태안반도이며, 동해안 북방한계지역은 강원도 지역으로 확인되었고, 번행초의 서해안 북방한계지역은 전라남도이고 동해안 북방한계지역은 경상북도 울릉군 지역이었다. 2. 자생지가 해안 200 m 이내 모래점토와 점토흙이 혼합된 지역에서 직사광선은 받는 우점식생이 없는 것을 볼 때 재배 지역은 해안지역과 인접한 지역은 재배 가능지역으로 판단된다. 3. 생리활성효능을 RAW 264.7 세포와 HL-60 세포를 이용하여 확인한 결과 유효한 항염효능은 확인하지 못하였고, 항암활성을 검정한 결과 순비기나무, 번행초는 억제활성이 없었고, 갯방풍은 100μg/ml에서 60%, 200μg/ml에서 72%의 증식억제 효과를 확인하였다.

Curcumin, a polyphenolic antioxidant purified from turmeric, has been known to possess various biological activities such as anti-oxidative, anti-inflammatory and anti-cancer effects. In this study, we have explored anti-inflammatory effect of curcumin using Gram (-) bacterium-derived endotoxin (lipopolysaccharide: LPS) and macrophage cell line RAW264.7. Curcumin suppressed NO production in LPS-activated RAW264.7 cells in a dose-dependent manner, Curcumin also blocked the activation of NF-kB but not AP-1 according to luciferase assay. Furthermore, this compound suppressed the phosphorylation of a series of intracellular signaling components such as Src, JAK-2, Akt, IKK and IkBα under LPS stimulation in a time dependent manner, Therefore, our data suggest that curcumin was able to protect the host from Gram(-) bacterial-infection-mediated inflammatory symptoms.

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects of the ethanol extracts of Rubus coreanus Miq. (ERC) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis factor-α (TNF-α), and Interferon-γ (IFN-γ) production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased IFN-γ and TNF-α production. Consistent with these results, the protein level of inducible Nitric Oxide Synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by ethanol extracts of ERC in a dose-dependent manner. These results suggest that ERC may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

Naturally occurring substances are important biomedical resources with low toxicity and ethnopharmacology-based efficacy. Four out of 45 extracts (Celastrus orbiculatus, Cercis chinensis, Stephanadra incisa, and Weigela subsessilis) prepared from the bark of Korea Forest plants exhibited more than 50% of inhibition on TNF-α production in lipopolysaccharide (LPS)-activated RAW264.7 cells at 100 μg/ml. In particular, potential inhibitory components of 4 extracts showed more than 50% inhibition seemed to be concentrated in methylene chloride (MC) fraction from C. orbiculatus, in ethyl acetate (EtOAc) fraction from C. chinensis and in hexane (Hx) fraction from S. incisa, whereas inhibitory activities of W. subssilis were broadly seen in non-polar solvent fractions such as Hx, MC and EtOAc. Therefore, our results suggest that extracts from C. orbiculatus, C. chinensis, S. incisa and W. subsessilis may be developed as a therapeutic remedy against TNF-α-mediated diseases such as rheumatoid arthritis or further fractionated to isolate active components having antiTNF-α inhibitory activity.