본 연구에서는 하수오를 이용하여 50% 에탄올 추출물과 에틸아세테이트 분획물을 제조하고, 추출물 및 분획물의 항산화, 세포 보호 및 항균 효능을 평가하여 기능성 소재로서의 가능성을 확인하였다. 또한, 하수오에 주요하게 존재하는 성분의 활성도 검증하였다. HPLC-DAD, LC-EIS-MS를 통해 분석한 결과, 하수오의 주성분은 2, 3, 5, 4'-tetrahydroxystilbene 2-O-β-D-glucoside (THSG)이었다. 페놀류 및 THSG 함량은 에틸아세테이트 분획물이 에탄올 추출물 보다 각각 11.1 및 3.0배 높게 나타났다. DPPH 자유 라디칼 및 Fe3+-EDTA/H2O2 시스템에서 생성된 ROS에 대한 소거 활성 평가에서, 에틸 아세테이트 분획물은 에탄올 추출물 보다 뛰어난 소거 활성을 나타냈다. 특히 ROS 소거 활성 평가에서 에틸 아세테이트 분획물과 THSG은 L-ascorbic acid와 동등한 소거 활성을 나타냈다. 이러한 결과를 바탕으로 수행된 광증감 반응으로 유도된 적혈구의 산화적 손상에 대한 세포 보호 활성은 에틸아세테이트 분획물, 에탄올 추출물, THSG 순서로 나타났으며, 모든 실험군이 양성 대조군으로 사용한 (+)-α-tocopherol보다 우수한 활성을 나타냄을 확인하였다. 항균 활성 평가는 S. aureus, E. coli, P. aeruginosa, C. albicans 균주를 대상으로, disc diffusion assay와 broth microdilution assay를 이용하여 수행하였다. 그 결과 추출물, 분획물 및 THSG 모두 모든 균주에 대해 항균 활성을 나타냈으며, 특히 그람 양성균인 S. aureus에 대해 methyl paraben보다 우수한 항균력을 나타냄을 확인하였다. 본 연구의 결과는 하수오가 항산화, 세포 보호 및 항균력에 관한 천연 소재로의 활용될 수 있는 가능성을 시사한다.

본 연구진은 메틸말리올라이드가 인간 피부 세포주인 HaCaT 세포에서 NRF2를 유도하고 이를 통하여 항산화 효과를 나타내는지 알아보았다. MTT assay를 통하여 메틸말리올라이드는 10 μ M 농도에서 24 h 동안 HaCaT 세포에 처리하여도 HaCaT 세포 독성을 나타내지 않는 것을 확인하였다. 본 연구진이 구축한 HaCaT-ARE-GFP-luciferase 세포에 메틸말리올라이드를 처리하고 루시퍼라아제 활성을 측정한 결과 메틸말리올라이드는 양성 대조군인 레스베라트롤보다 ARE 결합에 따른 루시퍼라아제 활성을 더욱 강하게 증가시키는 것을 확인하였다. 또한 HaCaT 세포에 메틸말리올라이드 처리 시 NRF2에 의하여 유도되는 항산화 단백질인 HO-1과 NQO1 mRNA 및 단백질 발현이 통계적으로 유의하게 증가하였다. 마지막으로 메틸말리올라이드는 HaCaT 세포에서 TPA에 의해서 유도된 DNA 및 지질의 산화를 강력하게 억제하였다. 결론적으로 본 연구는 메틸말리올라이드가 NRF2 유도를 통하여 피부 산화적 스트레스를 억제하며 이는 메틸말리올라이드가 신규 항산화 화장품의 소재로 적합하다는 것을 시사한다.

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: For stable induction of tetraploidy in Fallopia multiflora Haraldson, colchicine was treated to establish the condition of induction and investigated the morphological and cytogenetic traits of the tetraploid plants obtained compared to those of diploid ones. Methods and Results: For the induction of tetraploidy, F. multiflora plants were soaked in aqueous solutions of colchicine at various concentration (0.1, 0.5, and 1.0%). After this, 2% dimethyl sulfoxide (DMSO) was added at room temperature on a shaker set at 150 rpm for periods of 12, 24, and 48 h. The induction rate of tetraploids appeared to be the highest in plants treated with 0.5% colchicine for 24 h. As the colchicine concentration and soaking time increased above these levels, the growing tip of the roots did not develop and they began to rot. When compared to diploid plants, tetraploids differed greatly in various characteristics, including the sizes and shapes of the leaves, fruits, flowers and roots. The induced tetraploid F. multiflora had larger guard cells, and chloroplasts, increased number of chloroplast in the guard cells and decreased stomatal densities. Conclusions: When colchicine induced plants for tetraploid, it can be distinguished from diploids, in various characteristics such as morphological changes as stomatal size, number of chloroplasts per guard cell, number of chromosomes and flow cytometry. Therefore, it proved that these methods are suitable, quick and easy methods for the identification of the ploidy level of F. multiflora.

Background : Dry eyes are caused by highly increased osmolarity of tear film, inflammation, and apoptosis of the ocular surface. Polygonum cuspidatum is a herbaceous perennial plant of the genus Polygonum found in Asia and North America. However, the effects of P. cuspidatum aqueous extract (PCE) on hyperosmolarity-induced inflammation and apoptosis in human corneal epithelial cells have not been examined. Methods and Results : Hyperosmotic media induced human corneal epithelial cell (HCEC) cytotoxicity though increased inflammation, apoptosis, and oxidative stress. PCE treatment significantly inhibited expression of cyclooxygenase-2 and inflammatory cytokines (interleukin-6 and tumor necrosis factor-α), and activation of NF-κB p65 in hyperosmolar stress-induced HCECs. In addition, Hyperosmolarity-induced increase in BAX expression and activation of cleaved poly (ADP-ribose) polymerase and caspase 3 were attenuated in a concentration-dependent manner by PCE. PCE treatment restored anti-oxidative proteins such as Heme oxygenase-1 (HO-1), Superoxide dismutase-1 (SOD-1), and Glutathione peroxidase (GPx) in hyperosmolar stress-induced HCECs. Conclusion : PCE protected against hyperosmolar stress-induced inflammation, apoptosis, and oxidation by inhibiting expression of COX-2, BAX, MMP9; activation of NF-κB, caspase 3, and PARP; and increasing expression of MUC4 and anti-oxidative proteins. Overall, our data provide insight into the protective effects of PCE as a candidate for eye health.

Background : Rehmannia glutinosa (RG) has been utilized as a traditional medicine in Asia. However, the development of varieties is limited and the climate is changing gradually. Therefore, it is required to develop a superior lineage suitable for this. So we have secured several species, and it is necessary to confirm the cytotoxicity of various kinds of cells for its safety and to secure safety for further utilization. Methods and Results : 11 cultivars and 21 lineages of RG were collected from Rural Development Administration (RDA) at Eumseong of Chungcheongbuk-do and national farmhouse. They were cultivated in test-research farm in RDA and used as materials. Human (THP-1 cell, human leukemia monocytic cell line) and rodent-derived immune cells (RAW264.7, murine monocyte/macrophage cell line) and hepatocytes (HepG2, human liver cell line) were used to assess cytotoxicity. Cytotoxicity was determined by using MTT assay. As a result of evaluation of cytoxicity, 11 cultivars and 21 lineages of RG were not shown cytotoxicity range from 250 - 1,000 ㎍/㎖ concentration in THP-1 cell, RAW264.7 cell and HepG2 cell. Conclusion : Development of RG with superior lineage suitable for changing climate is required. We selected a good lineage (21 ea), and result of the cytotoxicity evaluation from low to high concentrations in immune- and liver-derived cells, there was no toxicity at all. Therefore, if these excellent lineages are distributed to farmers, they can help farmers. And it can help research on immunity and liver function in the future.

Background : As a part of ongoing research to elucidate and characterize anti-cancer nutraceuticals, extracts from many kinds of medicinal plants were tested for their ability to cytotoxicity on cancer cells so far. Datura stramonium is one of the plants known to contain various alkaloids such as hyoscyamine, scopolamine, atropine etc. Traditionally, it has been used as an analgesic, antispasmodic, and central nervous stimulant. Leaves are also known to be effective against asthma, cough, and chronic bronchitis. In this study, cytotoxicity of extracts from D. stramonium on human cancer cell lines A549 (lung), and HepG2 (liver) were evaluated and compared. Methods and Results : The extract was diluted with DMSO in the form of 10, 50, 250, 1,000 μg/ml for final concentration series respectively. The cell viabilities were examined by MTT assay. On HepG2 cell line, extracts of D. stramonium showed dramatic dose-dependent cytotoxicity on 10, 50, 250, 1,000 ㎍/㎖ concentrations series as 88.16%, 78.55%, 55.57%, 23.06% cell viability respectively. On A549 cell line, likewise, same concentration series showed a dose-dependent cytotoxic effect as 96.49%, 96.12%, 68.54%, 20.26% cell viability respectively. On A549 cell line, there was no difference in effect between 10 ㎍/㎖ and 50 ㎍/㎖. Conclusion : Above results, high concentrations of extracts are effective on two cancer cell lines. These results are expected to be used on further studies about the anticancer activity of D. stramonium as basic data. In order to confirm the anticancer effect of D. stramonium, it is anticipated that additional tests will be required to confirm the apoptosis assay and related protein expression.

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.

Background : Obesity is a type of metabolic diseases caused by unbalanced in take and consumption of calories. 3T3-L1 is differentiated into adipocytes by various hormones and transcription factors and accumulates intracellular lipid. Therefore, it is important to inhibit the adipocyte differentiation precess for effective obesity inhibition. Aster yomena and Aster glehui are medicinal plant of Compositea family that grows widely in Korea. Aster genus plants have been used to treat snakebite wound or bruises in oriental. The aim of this study was comparison of inhibitory effect oxidation and adipocyte differentiation with Arial parts of A. yomena (AY) and A. glehni (AG). Methods and Results : AY and AG were cultivated from Pyeongchang in Korea, 2018. AY and AG were extracted by 70% ethanol (-E) and water (-W) at room temperature. AG-W has higher phenolic content (6.92 ± 0.23) and AG-E has higher flavonoid content (8.22 ± 0.19) than other extracts. AG-E has higher radical scavenging activity on DPPH and ABTS assay (IC50 value; 104.88 ± 10.50 and 30.06 ± 0.27). In cytotoxity assay, all extracts concentrations of lower 100 ㎍/㎖ were nontoxic to the cells and can be applied for the next assay. The anti-adipogenic effect of extracts were determined in 3T3-L1 cell by Oli Red-O (ORO) staining. The lipid diplot stained with Oil red O was dissolved to determine by microplate-reader. AG-W significantly reduced the adipocyte differentiation of 3T3-L1 cells (70.49%) compared with other extracts (AG-E, AY-W and AY-E). Conclusion : Theses results reveal that the water extract of AG has utility as a functional food material for preventing obesity.

Background : Osteoclasts are differentiated from the monocytes/macrophages of hematopoietic cells, that excessive activities of bone-resorbing giant cells leads to pathological bone diseases such as osteoporosis (contained rheumatoid arthritis and autoimmune arthritis). Therefore, it is very important to suppress loss of bone mass by deactivation of osteoclast differentiation. In this context, we evaluated for the effects of black ginseng (BG) extract on TRAP activity, proliferation and differentiation in RANKL-induced osteoclastic RAW264.7 cells. Methods and Results : The aim of this study is to figure out the potential anti-osteoporosis effects and the underlying mechanism of BG extract in RANKL-induced osteoclastic RAW264.7 cells. The ginsenoside Rg3, Rg5, Rk1 and Rh4 content of BG was increased more than Red ginseng (RG). The extracts of BG markedly reduced the activity of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from osteoclastic RAW264.7 cells, without cytotoxicity. BG clearly inhibited RANKL-induced osteoclast differentiation by decreased calcitonin and TRAP (p < 0.01). Furthermore, ginseonside Rg5 and Rk1 significantly inhibited TRAP activity in formation of osteoclastic differentiation (p < 0.01). It is also found that Ginseonside Rg5 and Rk1 mixture more inhibits osteoclast differentiation activity. Conclusion : Our results suggest that Black ginseng extract has an anti-osteoporosis effects in bone disease when administered as a food supplement and has potential as a therapeutic agent for osteoporosis.

Background : Gastrodia elata (GE) is a perennial herb that belongs to the orchidaceae and is used as a medicinal or food material. Known pharmacological agents include gastrodin and 4-hydroxybenzyl alcohol. It is used as medicinal herb that is traditionally used for headache, migraine, dizziness, epilepsy and infant seizures. It is used for medicinal herbs such as sedation, hypnosis, epilepsy treatment, anticonvulsant, antidepressant, neuroprotection, antipsychotic, anticonvulsant, Antioxidant, memory improvement, anti-aging, antiviral, anti-tumor. The purpose of this study was to find the extraction method with the highest oxidative stress inhibition and to optimize the pharmacological effect of the extract. Methods and Results : GE was freeze-dried to obtain 5 g, and then extracted into 50 ㎖ of water. Extraction temperature was 0, 30, 60 and 90℃ for 20, 40, 60 and 120 min, respectively. After centrifugation, the mixture was filtered through a 0.45 ㎛ filter. ABTS scavenging ability, DPPH scavenging ability, total phenol content, neuronal cell line (PC12) cytotoxicity, and oxidative stress scavenging activity in neurons were measured by this extract. ABTS scavenging ability, DPPH scavenging ability and total phenol content increased with increasing temperature and extraction time. However, at 60℃ and 90℃ extraction temperature, there was no significant difference. The cytotoxicity of 2 ㎎/㎖ of GE extract was significantly increased in the extract group of 90℃ after 20 hours. Conclusion : From the above results, the water extraction conditions to optimize the pharmacological activity of GE were 120 minutes at 60℃ or less.

This study was designed to compare re-epithelization rate/feature of conjunctival epithelial cell on amniotic membrane and anatomical difference between normal amniotic membrane and de-epithelial amniotic membrane. Human conjunctival tissue obtained while cataract operation were used in this study. Primary cultured human conjunctival epithelial cell and fibroblast were incubated on intact amniotic membrane and de-epithelialized amniotic membrane. After incubation for 3, 5 and 7 days, re-epithelization rate was analyzed using cell tracker and microscopic feature was examined using methylene blue and hematoxylin- eosin stain. For cell marker analysis, cytokeratin 14 and vimentin antibody immunofluorescence stain were used. Clearance rate of epithelial cells on amniotic membrane using trypsin was better than using 25% alcohol. The results of reepithelization rate using cell tracker, HaCaT, conjunctival firbroblast, and epithelial cell were rapidly incubated on deepithelialized amniotic membrane. Wound healing and re-epithelization were more rapidly induced on de-epithelialized amniotic membrane on permanent amniotic membrane graft.

Background : Ginseng seeds are harvested in immature stages and take 3 - 4 month to be germinated following cold stratification. Improvement of germination speed can be a valuable traits to be characterized for economically important medicinal plant. Modulation of cell wall compositions is also considered as another agricultural trait when the crops can be processed for foods and forages. Here we show the functional characterization of ginseng pPLAIIIβ focused on its possible values on germination and modulation of cell wall compositions. Methods and Results : Patatin-related phospholipase AIII family genes were identified from ginseng EST clones and further confirmed by PCR. Clone showing the highest homology with pPLAIIIβ was overexpressed in model plant Arabidopsis, and it displayed dwarf plants. qPCR analysis showed that pPLAIIIβ expressed in all organs of 2-years-old ginseng. Overexpression of ginseng pPLAIIIβ also displayed apparently enlarged seeds as well as altered germination speed in early stage compared to the control. Chemical stainings and direct quantification of lignin and cellulose were performed to understand the link of cell elongation patterns and cell wall compositions. Conclusion : Overexpression of ginseng pPLAIIIβ inhibited longitudinal cell elongation in all tested organs except seeds which is enlarged in both directions than the control. Shorter root length is related with auxin responsive genes and its dwarf morphological phenotype is resulted in altered cell wall compositions.

Background : Turmeric (Curcuma longa L.) and black pepper (Piper nigrum L.) have traditionally been used as Asian medicinal and culinary herb. Curcumin, a major compound of turmeric, has been known to have antitumor activity. However, curcumin is bioavailable because it is rapidly metabolized and released from the body. Therefore, the addition of adjuvants such as piperine, a potent inhibitor of drug metabolism, is one of the ways to increase the bioavailability of curcumin. Methods and Results : The yields of turmeric and black pepper ethanolic extracts (TM and BP) are 18.2 and 8.2% (w/w), respectively. The EC50 values of A549 and NCI-H292 cells exposed to TM were 77.8 ㎍/㎖ and 92.0 ㎍/㎖, respectively. No significant cytotoxicity was observed up to the 400 ㎍/㎖ in the A549 and NCI-H292 exposed to BP. Based on the central composite design, the co-treatment of TM and BP enhanced the cytotoxicity of A549 and NCI-H292 cells. The optimal combination concentration (optimal EC50 value) of TM and BP calculated by the response surface methodology assay were 48.5 and 241.7 ㎍/㎖. The conbination index assay confirmed that the cytotoxic effect at optimal combinatino concentration was due to the synergistic effect. Conclusion : We hypothesized that co-treatment of TM and BP enhanced cytotoxicity more than single treatment of TM against lung cancer cells, and cell death at this time may synergetic cytotoxicity effects associated with curcumin metabolism.

천연물 유래 단일 성분의 지방세포로의 분화 및 지방축적 억제효과에 대한 잠재성을 탐색하기 위해 본 연구에서는 화살나무의 껍질에서 분리 정제한 FG가 3T3-L1 지방전구세포에 있어서 지방세포로의 분화 및 지방구 축적에 미치는 영향에 대해 평가하고, 관련 메카니즘에 대해 검토하였다. FG는 분화용 배지에 포함된 분화 촉진인자들의 자극에 의한 지방생성에 있어 주요한 전사인자인 PPARγ와 그 표적 단백질인 FABP4의 발현을 억제함으로써 지방세포 분화 및 지방구 축적을 억제하는 것으로 나타났다. 이상의 결과로부터 FG가 비만이나 비만과 관련된 당뇨병 등을 예방, 개선하는데 유용한 물질로 사용될 가능성이 있을 것으로 생각된다.

화장품에 방부제(살균/보존제)로 사용되는 1, 2-hexanediol (HD)로 인한 부작용을 극복하기 위하여, Escherichia coli (E. coli)의 β-galactosidase (β-gal)를 이용하여 transgalactosylation 반응으로 1, 2-hexanediol galactoside (HD-Gal)를 합성하였다. 본 연구에서는 합성된 HD-Gal의 인간 피부세포에 대한 독성이 어느 정도인지를 HD와 비교하여 관찰하였다. HD-Gal과 HD의 세포독성은 인간 피부각질형성세포 (HaCaT cell line)에 HD와 HD-Gal을 처리한 후, cell proliferation assay를 이용하여 비교 분석하였다. 또한 이때, 위상차 현미경으로 HD-Gal과 HD로 처리한 세포의 상태를 비교 관찰하였다. 그 결과, HD-Gal은 42.2 mM에서 211 mM의 농도 범위에서 세포독성이 관찰되지 않았으며, 현미경 관찰에서도 큰 변화를 관찰할 수 없었다. 그러나, HD의 경우에는 저농도에서(42.2 mM and 84.4 mM)는 세포독성이 관찰되지 않았으나, 고농도 (168.8 and 211 mM)에서 매우 높은 세포독성을 나타내었고, 현미경 관찰에서는 고농도에서는 물론이고, 세포 독성이 관찰되지 않은 HD의 저농도에서도 세포모양과 세포 수에서의 변화가 관찰되었다. 앞으로 세포독성이 감소된 HD-Gal이 HD의 대체제로서 안전, 건강 및 웰빙 개념의 새로운 용도로 개발될 수 있을 것으로 생각된다.

A diarylheptanoid, (5S)-1,7-bis-(3,4-dihydroxyphenyl)-5- hydroxyheptane-3-one-5-O-β-D-xylopyranoside, named oregonin (1), was isolated from the of Alnus japonica (A. japonica), which is a species of the genus Betulaceae, growing throughout Korea, Japan and China. The structure was elucidated by various spectroscopic methods including negative and positive LC/MS, 1H-NMR, and 13C-NMR techniques or by comparison with authentic samples. In order to evaluate the anti-oxidative activities of oregonin (1) isolated from A. japonica, 1,1-diphenyl-2-picryhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis[3-ethylbenzothiazoline-6-sulphonic acid] (ABTS) radical scavenging activity were measured in vitro. Oregonin from A. japonica exhibited potent DPPH and ABTS radical scavenging activities. A. japonica shows not only 1,1-diphenyl-2-picryhydrazyl (DPPH) radical scavenging activity and ABTS radical scavenging activity, but also apoptosis modulative effects. The present results indicate that A. japonica could be a hair-growth-promoting agent for cosmetic products.

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.

대기 오염은 피부의 산화적 손상, 염증 및 노화를 일으킬 수 있다. 레스베라트롤은 폴리페놀 화합물의 일종으로 항산화, 항염증, 멜라닌 생성 억제 작용 등 다양한 생물학적 활성이 있는 한편 열과 빛에 약한 단점이 있다. 레스베라트릴 트라이아세테이트(RTA)는 레스베라트롤에 비해 안정하고, 피부 안전성과 미백 효능이 보고된 화장품 신소재이다. 본 연구의 목적은 직경 10 μ m 미만 대기 미립자 물질(PM10)에 노출된 인간 표피 각질형성세포(HEK)의 염증 반응에 대한 레스베라트롤과 RTA의 영향을 조사하기 위한 것이다. 배양된 HEK세 포를 레스베라트롤과 RTA의 유무 조건에서 PM10에 노출시키고, 세포 생존율, 반응성 산소종(ROS)의 생성 및 염증성 사이토카인의 발현을 분석하였다. PM10을 처리하였을 때 세포 생존율이 감소하였고 종양괴사인자- α(TNF-α), 인터루킨-1β(IL-1β), 인터루킨-6(IL-6) 및 인터루킨-8(IL-8)의 발현이 증가하였다. 레 스베라트롤과 RTA는 PM10으로 유도된 세포의 사멸과 ROS 생성을 경감시켰다. PM10에 의해 증가되는 여러 염증성 사이토카인의 발현은 레스베라트롤과 RTA에 의해 경감되거나(IL-6), 증진되거나(IL-1β), 변화하지 않았다(TNF-α 및 IL-8). PM10에 의해 유도된 IL-6단백질의 발현이 레스베라트롤과 RTA에 의해 감소되 었다. 본 연구의 결과는 레스베라트롤과 RTA가 대기 미립자 물질에 노출된 피부의 세포 손상과 염증 반응을 조절하는 작용이 있음을 시사한다.

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.