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.

Background : In ancient, roots of Rumex crispus, called wooi-daehwang, were used for various symptoms and diseases like cough, phlegm, bronchitis and hepatitis, caused inflammatory. As a part of ongoing research to elucidate and characterize anti-inflammatory nutraceuticals, solvent-partitioned fractions from R. crispus root were tested for their ability to suppress inflammation. In this study, NO synthesis inhibitory activity of solvent-partitioned fractions from R. crispus root on LPS-stimulated RAW264.7 mouse macrophages was evaluated. Methods and Results : The EtOH extracts were suspended in water. The aqueous layer was further partitioned in diethylether, ethylacetate and n-butanol, sequentially. RAW264.7 cells were seeded onto 96-well plates, and cells were allowed to adhere for 6 h and then were pretreated with the R. crispus root extracts for 24 h. Cellular nitric oxide (NO) production was stimulated by adding lipopolysaccharide (LPS). Absorbance was measured at 520 ㎚ by microplate reader. NO synthesis inhibitory activity potential of these fractions was evaluated by assessing NO production by LPS-stimulated RAW264.7 cells in the presence and absence of the solvent-partitioned R. crispus root fractions. NO synthesis inhibitory activity of diethylether fraction diluted in 50 ㎍/㎖, 25 ㎍/㎖, 12.5 ㎍/㎖, 6.25 ㎍/㎖, 3.125 ㎍/㎖ was 79.2%, 70.9%, 59.5%, 16.1%, and 11.8%, respectively. And NO synthesis inhibitory activity range of another fractions, EtOAc, n-BuOH and aqueous layer, were 0 - 30.2%, 0 - 20.1% and 3.8 - 22.4%, respectively. Conclusion : From the above results, it showed that diethylether fraction have strong NO synthesis inhibitory activity, it was suggested that R. crispus root have NO synthesis inhibitory effects. R. crispus root possesses anthraquinones, such as chrysophanol, parietin, and anthrones etc. According to previous studies, R. crispus semen extract has analgesic and hepatoprotective effect as anti-inflammatory, and extract of R. napalensis has cyclooxygenase (COX)-2, COX-1 inhibitory and free radical scavenging effect. Our present study has shown that R. crispus root extracts anti-inflammatory effects probably by suppressing iNOS expressions, and resulting in the inhibition of NO synthesis.

만성적인 염증은 낭포성 섬유증, 암, 치매, 아토피성 질환, 비만 등과 같은 염증성 질환의 원인이 된다. 또한 염증의 발생단계에 관여하는 일부 신호물질은 피부조직의 손상과 노화에도 영향을 주는 것으로 알려져 있기 때문에 염증발생 매커니즘을 조절하기 위한 연구가 활발하게 이루어지고 있다. 최근에는 염증반응을 억제하거나 예방하기 위해, 몇몇 식물로부터 항염증 소재를 개발하려는 연구들이 이루어지고 있다. 특히 Stevia rebaudiana는 특유의 풍미를 가지는 천연감미료 스테비올배당체(steviol glycoside, SG)를 생성하는데, 일부 SG 에 대한 연구를 통해 염증억제 활성이 있는 것으로 알려져 있다. 본 연구에서는 기존연구를 통해 항염증 효능이 있는 것으로 확인된 스테비오사이드, 리버디오사이드 A, 스테비올 이외에도 항염증 소재로 활용될 가능성이 있는 SG가 더 존재할 것으로 예상하였다. 이를 확인하기 위하여 우리는 S. rebaudiana에서 얻은 SG의 nitric oxide (NO) 생성억제활성을 RAW 264.7 세포주를 대상으로 스크리닝 하였다. 그 결과 steviol β-glucopyranosyl ester (SGE)가 동일한 농도 조건의 SG 중에서 가장 높은 억제활성을 보여주었다. 또한, interleukin-1α (IL-1 α), interleukin-1β (IL-1β), cyclooxygenase-2 (COX-2), nuclear factor kappa-light chain-enhancer of activated B cells (NF-κB), inducible nitric oxide synthase (iNOS)와 같은 염증관련 인자의 mRNA 발현량 또한 농도의존적으로 감소시키는 것으로 확인되었다. 이러한 연구결과를 통해 SGE는 마우스 대식세포인 RAW 264.7 세포에서 항염증 활성 및 NO 생성 억제 효과가 있음을 확인하였다. 이를 통하여 SGE가 항염증 소재로 활용될 잠재성이 있음을 확인하였다.

Background: The flowering plant Hippophae rhamnoides L. has been used for many studies on fruit or leaf extracts. This study was conducted to investigate the development of a new cosmetic material from H. rhamnoides fruits and leaves that have by antioxidant, anti-inflammatory and wrinkle improvement activities. Methods and Results: The antioxidant abilities of H. rhamnoides extracts, including of a water-soluble fruit powder (FW), a fatsoluble fruit powder (FF), a supercritical extract of fruit by-product (BS), and a mixture of leaf and fruit (MIX), were investigated in vitro. A DPPH radical assay for antioxidant activity was performed for these fractions alongside assay to evaluate the total phenolic and flavonoid content (TPC and TFC). As expected, the MIX had the highest DPPH radical scavenging activity (RC50 = 10.27㎍/㎖), and the TPC and TFC also were highest in MIX (225.7 ㎎·GAE/g, and 25.18 ㎎·QE/g, respectively). Nitric oxide (NO) production in LPS-induced RAW264.7 cells was estimated and the results indicated an over 75% decrease of NO production in FF and MIX. In other assays, the highest elastase inhibitory activity was found in FW. Conclusions: These results revealed that H. rhamnoides extracts have a high potential for antioxidant, anti-inflammatory and antiwrinkle activities. H. rhamnoides products are suggested to be applied as the functional materials of cosmetic ingredients.

Background: Inula japonica Thunb. is a plant belonging to the family compositae. Inulae flos (flower of I. britannica var. chinensis Regal.) is the dried flower of I. japonica Thunb. and contains various flavonoids (patulitrin, nepitrin and kaempferol), which have been utilized in traditional oriental medicine to treat nausea, phlegm, and coughs. However, ethanol extract of I. britannica (IJE) has not been previously studied for its use in cancer treatment, and its effects on oxidative stress, or inflammation. Thus, the present study investigated the anti-oxidant, anti-inflammatory, and anti-colorectal cancer effects of IJE using RAW264.7 and HCT- 116 cells, which are human colorectal cancer cell line. Methods and Results: IJE contained flavonoids (80.95 ± 5.3 ㎎/g) and polyphenols (310.53 ± 10.6 ㎎/g). Moreover, it reduced lipopolysaccharide (LPS)-induced nitric oxide (NO) production and H2O2-induced oxidative stress by decreasing reactive oxygen species (ROS) levels. Additionally, the 500 ㎍/㎖ IJE treatment increased caspase-3 activity and apoptotic cell death in HCT-116 cells. Conclusions: These results demonstrate that the anti-cancer effect of IJE against human colorectal cancer cells involves caspase-3 activation and apoptotic cell death. IJE also inhibited LPS-induced NO production, and H2O2-induced oxidative stress in RAW264.7 cells. However, further studies are required to explore how IJE treatment regulates signal transduction in NO and ROS production.

해방풍이 가지는 건강 기능성 소재로서의 가치를 확인하고자, 해방풍 부위별 용매추출물의 항산화 활성 및 면역조절 효과를 조사하였다. 해방풍 부위별 용매추출물의 추출수율은 10.73-30.57이며, 총 폴리페놀 및 총 플라보노이드 함량은 해방풍 잎 70% 에탄올 추출물(EL)에서 각각 10.79 g/100 g 및 2.01 g/100 g으로 가장 높은 값을 나타내었다. DPPH radical 및 ABTS radical 소거활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 12.90-84.70% 및 11.78-57.64%로 가장 우수한 radical 소거활성을 나타내었으며, superoxide radical 소거활성 및 FRAP 활성은 해방풍 잎 70% 에탄올 추출물(EL)이 100-1,000 μg/mL 농도에서 각각 49.91-84.05% 및 255.38-975.28 μM로 가장 우수한 활성을 나타내었다. 대식세포주인 RAW 264.7 세포에 해방풍 부위별 용매추출물을 처리하였을 때 nitric oxide 생성 억제능을 확인하기 위해 분석한 결과, 추출용매에 따른 nitric oxide 생성량은 70% 에탄올 및 70% 메탄올 추출물에서 가장 낮은 생성량을 보였으며, 해방풍 부위별에 따른 nitric oxide 생성량은 뿌리, 잎 및 씨앗 순으로 낮았다. 그 중에서 해방풍 뿌리 70% 에탄올 추출물(ER)에서 가장 낮은 nitric oxide 생성량을 보여 우수한 nitric oxide 억제 효과를 확인하였으며, 이를 이용하여 기능성 소재, 미용식품 및 화장품 제조용 원료 등 다양하게 활용 가능할 것으로 사료된다.

Background: To date, the anti-wrinkle efficacy of phellodendri cortex has not been defined. In this study, we investigated the nitric oxide (NO) production and elastase inhibitory activities of 80% methanol extract of Phellodendri cortex and its ethyl acetate fraction. Methods and Results: We prepared 80% methanol extract, and its fractions from phellodendri cortex. The treatment of RAW 264.7 cell with 25㎍/㎖ 80% methanol extract and ethyl acetate fraction resulted in no toxicity. We conducted assays of nitric oxide (NO) production and elastase inhibition. In the NO production assay, the ethyl acetate fraction showed an inhibitory effect approximately 17 times stronger than the 80% methanol extract. In elastase inhibitory assay, the ethyl acetate fraction also showed a stronger effect than the 80% methanol extract. In order to standardize the extract and fraction, we used TLC to separate the extract and observed the plate under UV light. We confirmed that the known pharmacological ingredients berberine, and palmatine in the 80% methanol extract and the ethyl acetate fraction. Conclusions: These results indicated that phellodendri cortex extract and its ethyl acetate fraction produced strong inhibitory effect on elastase and NO production.

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 : 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.

Background: In this study, we investigated the antibacterial and nitric oxide (NO) production inhibitory activities of 75% ethanol extract of Prunus sargentii branches and its fractions against acne pathogens. Methods and Results: The antibacterial activity against acne causing pathogens was determined using the disc diffusion assay. The ethyl acetate fraction showed higher activities against Propionibacterium acnes, Staphylococcus aureus and Staphylococcus epidermidis than those shown by other fractions. In the DPPH radical and NO scavenging assays, the butanol fraction showed strong DPPH radical and NO scavenging abilities. These activities were related to the total polyphenol and flavonoid contents of butanol fraction. On the other hand, the chloroform and ethyl acetate fractions exhibited the highest NO production inhibitory activity in Lipopolysaccharide (LPS)-stimulated Raw 264.7 cells compared to those exhibited by other fractions. Conclusions: The extract and its ethyl acetate fraction from the branches of P. sargentii exhibited antibacterial activity and could be used as functional materials in antimicrobial related fields. Moreover, the chloroform and ethyl acetate fractions are potential antiinflammatory agents and butanol fraction acts as an effective radical scavenger.

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.

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.