Oxidative stress caused by reactive oxygen species (ROS) is a key mechanism of skin aging, and the use of antioxidants is an effective strategy to prevent the symptoms associated with ROS-induced skin aging. The components rich in polyphenolic compounds with antioxidative activity were identified by fractionating an aqueous ethanolic extract of Gryllus bimaculatus (AE-GBE) using solvent using hexane, chloroform, ethyl acetate, butanol, and water. Their ability to migrate H2O2-induced oxidative stress in human dermal fibroblasts (HDFs) was then evaluated. The butanol fraction of AE-GBE had the highest polyphenol content and antioxidant effect, followed by the ethyl acetate and water fractions, suggesting that the likely antioxidant components are polar components. Furthermore, the butanol, ethyl acetate, and water fractions effectively reduced intracellular ROS production and DNA damage in HDF cells caused by H2O2. Overall, these findings suggest that the butanol fraction of AE-GBE shows promise as a natural insect-derived antioxidant material, capable of suppressing oxidative stress by showing a stronger antioxidant effect under H2O2 stimulation than the other fractions.

본 연구에서는 주박의 기능성 화장품 소재로서의 활용 가능성을 평가하기 위해 분획물에 따 른 항산화 활성평가와 인간섬유아세포인 CCD-986sk 세포에서의 주름효능 평가를 진행하였다. ABTS+ radical 소거능 측정을 통해 항산화 활성을 확인한 결과, 주박 Ethyl Acetate 분획물 1,000 μg/ml 농도 에서 75.5%로 나타나 분획 용매 중 가장 높은 항산화 활성을 보였다. Elastase, Collagenase 저해 활성 측정을 통해 주름 개선 효과를 확인하였으며, 두 실험 결과 모두 주박 Ethyl Acetate 분획물이 1,000 μ g/ml 농도에서 가장 높은 효능을 나타내었다. UVB로 유도된 CCD-986sk 세포 내 pro-collagen type I 의 합성율을 측정한 결과 주박은 동일농도에서 20 μg/ml에서 Ethyl Acetate, Water, Acetonitrile, Hexan 분획물 순으로 높은 효능을 보였다. 콜라겐 분해효소인 MMP-1의 저해률을 측정한 결과 4가지 용매 분획몰은 20 μg/ml에서 모두 70% 이상의 효능을 나타내었다. Real time PCR 실험에서 pro-collagen type I, MMP-1 및 MMP-3의 mRNA 발현량을 측정한 결과 UVB 단독군 대비 Jubak 추출물을 함께 처리했을 때 pro-collagen type I의 단백질 발현량이 증가되었고, MMP-1과 MMP-3의 mRNA 발현량은 감소하는 것을 확인하였으며 대조군(EGCG) 다음으로 Ethyl Acetate 추출물이 주름 개선에 가장 효능이 우수하였다. 이상의 결과로 주박의 용매 분획물 중 Ethyl Acetate 분획물은 UVB 자극에 의한 광노화에 대한 항주름 효과가 있음을 확인하여 화장품의 천연물 소재로서의 이용이 기대된 다.

한국 자생식물인 기린초(Sedum kamtschaticum Fisch.)는 열수 추출 형태로 혈액 순환 개선과 항산화 및 항염증 효과를 내는 한약재로 사용되어왔다. 선행연구를 통해 기린초 여러 부위에서 우수한 항산화 효과를 확인하여, 본 연구에서는 기린초의 줄기(Stem)와 뿌리(Root) 부위를 각각 70% ethanol (SKS, SKR)로 추출한 후, n-hexane (SSH), ethyl acetate (SSE, SRE), chloroform (SSC, SRC) 및 water (SSW, SRW) 순서로 용매 극성별로 분획하여 화장품 소재로서 응용 가능성을 확인하였다. 각 분획물마다 총 폴리페놀 함량, 플라보노이드 함량 및 DPPH 라디칼 소거능을 확인한 결과 뿌리추출물(SKR)이 줄기추출물 (SKS)보다 우수한 함량과 효과를 나타냈고, 전체적으로 ethyl acetate 분획물(SSE, SRE)이 가장 효과가 우수했다. 또한, tyrosinase 활성 저해 효과도 ethyl acetate 분획물이 가장 우수하였고, 모든 분획물은 세포독성이 없는 10 μg/mL 농도에서 B16F10 멜라노마 세포에 처리했을 때 우수한 멜라닌 생성 저해력을 보였다. 70% ethanol 추출물(SKS, SKR)에 대해 HPLC 분석 시 gallic acid와 quercetin을 포함한 플라보노이드가 검출되었다. 본 연구에서는 기린초의 피부 항산화 효과와 미백효과를 나타내는 기능성화장품 천연소재로서 응용 가능성을 제시한다.

야생 잎새버섯 추출물을 이용하여 심혈관계 질환과 관련된 기능성 식품 개발을 위한 기초자료를 얻기 위해 준비한 물 추출물과 유기용매 분획물의 혈전용해활성과 트롬빈저해활성, acetylcholinesterase 저해활성, 항산화활성을 확인하였다. 10 mg/mL의 농도에서 물 추출물과 물 분획물은 각각 0.93 plasmin unit와 0.73 plasmin unit의 높은 혈전용해활성을 나타냈으며, 부탄올 분획물은 79.60%의 트롬빈저해활성을 나타냈다. 클로로포름 분획물은 85.88%의 높은 acetylcholinesterase 저해활성을 나타냈고, 물 추출물의 항산화 활성은 39.81%로 비교적 낮게 나타 냈다. 부분 정제된 효소를 포함하고 있는 물 분획물은 섬유소원의 Bβ chain를 모두 분해시켰지만, Aα chain은 느리게 분해시키고 γ chain 과는 반응하지 않았다. 실험 결과로부터 야생 잎새버섯 추출물은 높은 혈전용해활성과, 트롬빈저해활성, acetylcholinesterase 저해활성을 나타내 심혈관계 질환 예방을 위한 제약과 기능성식품 개발에 이용 가능할 것으로 기대된다.

Raphiolepis indica (R. indica) is one of evergreen shrubs belonging to the Rosaceae and is grown wildly in Jeju. This study was performed to evaluate the hepatoprotective effect of different fractions (n-hexane, dichloromethane, ethyl acetate, butanol, water) from R. indica. Anti-oxidative effects were determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity and total phenol contents. Hepatoprotective effect was identified by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay in Huh7 cells. Among various fractions, ethyl acetate and butanol fractions showed the lowest DPPH remained rate reaching approximately 78.7 and 65.5% at 400 μg/mL. Ethyl acetate and butanol fractions showed the total phenolic content at 164.5 and 137.3 mg GAE/g extract. The ethyl acetate and butanol fractions were resistant against oxidative stress in MTT assay and showed higher hepatoprotective effect than other fractions. Therefore, these results suggest that the ethyl acetate and butanol fractions of R. indica might have therapeutic value in liver damage.

This study was performed in order to analyze the fibrinolytic, thrombin inhibitory, anti-oxidative, acetylcholinesteraseinhibitory, and immuno-enhancing activities of the water extract and solvent fractions isolated from Grifola frondosa. Fibrinolyticactivity was analyzed using the fibrin plate method, and thrombin inhibitory activity was assayed using the substrate H-D-Phe-pip-arg-pna. Anti-oxidative activity was estimated using the DPPH assay, and AChE inhibitory activity was measured using thespectrophotometric method. Immuno-enhancing activity was examined using the nitric oxide (NO) production in RAW 264.7macrophage cells. Cell viability was determined using the MTS assay. Fibrinolytic activities were the highest in water extract (1.55plasmin units/mL) followed by water fraction (0.85 plasmin units/mL). The thrombin inhibitory activities of the water and ethylacetate fractions were determined to be 76.43% and 72.59%, respectively. The acetylcholinesterase inhibitory activities ofchloroform and hexane fractions exhibited values of 95.14% and 94.74%, respectively. The butanol fraction showed the highestanti-oxidative activity at 94.47%. Anti-proliferating activity against Raw 264.7 cells showed no cytotoxicity. The production of NOin Raw 264.7 cells increased up to 2-fold by adding the water fraction compared to the untreated control. These results suggestthat Grifola frondosa may serve as a useful functional food for the enhancement of immune function and the prevention andtherapy of cardiovascular diseases.

The purpose of this study is to determine the possibility of using Crataegi fructus as natural health food source. The research was conducted to determine the biofunctional activities of Crataegi fructus extract. Methanolic extract from Crataegi fructus was partitioned by using organic solvents, including n-hexane, ethyl acetate, n butanol, and water. Ethyl acetate soluble fraction was shown to have the strongest antioxidant activity (RC₅₀=4.39 ㎍/㎖) among the fractions. In the antimicrobial activity assays, ethyl acetate soluble fraction was effective in bacterial inhibition, against the cases of Escherichia coli and Klebsiella pneumonia, with minimum inhibitory concentrations in 125 ㎍/㎖. In the anticomplementary activity assays, water soluble fraction was the most effective exhibiting 18.4% inhibitory activity.

Twenty four fractions by solvent extraction and/or acid precipitation from fruit body and culture broth of Inonotus obliquus were prepared, and their inhibitory effect against acetylcholinesterase (AChE) was investigated. Among these fractions, acid (1 M HCl) precipitates from cell-free culture broth and fruit body exhibited the highest inhibitory effect on AChE in vitro. Acid precipitates inhibited AChE activity in a concentration-dependant manner and IC50 values of both acid precipitates were 0.53 mg/mL. The inhibition pattern was general non-competitive inhibition. The energetic parameters were also determined by dual substrate/temperature design. Both acid precipitates increased the values of Ea, ΔH, ΔG and ΔH* decreasing the value of ΔS for AChE. The results implied that the acid precipitates from I. obliquus increased the thermodynamic barrier, leading to the breakdown of ES complex and the formation of products as inhibitory mechanism.

Auricularia auricula-judae has long been used as food and traditional remedies in Asian countries such as Korea and China. In this study, we evaluated the in vitro anti-tumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae using various tumor cell lines. To do this, the mesh of Auricularia auricula-judae was mixed with 70% ethanol and heated at 1000C for 6 hrs and ethanol extract (ETOH) was collected. Ethanol extract was fractionated with dichloromethane (DCM), ethyl acetate, n-butanol and a water extract at room temperature as well as concentrated in a vacuum concentrator at a controlled temperature(<500C). The P388D1 macrophage and Sarcoma 180, human NSCLC NCI H358 (bronchioalveolar) and SNU1 cells (Gastric carcinoma) were cultured in RPMI. As the results, the cytotoxicity of the fractional extracts decreased significantly (P<0.05) in a dose-dependent manner. Dichloromethane extract (1 mg/ml) was the highest (P<0.05) in all experimental cell lines. There was also a significantly different sensitivity (P<0.05) among the P388D1, Sarcoma 180, NCI H358 and SNU1 cells for the fractional extracts. According to IC50 values, the most potent cytotoxic activity of dichloromethane fraction was found in Sarcoma 180 and NCI H358 cell lines. Butanol fraction appeared more cytotoxic to SNU1 cell line and water fraction had the highest cytotoxicity in P388D1 cell line. We did not find any significant difference between MTT and SRB assays in their ability to estimate cytotoxicity in all cell lines. Our findings suggest a potent antitumor activity of various fractions from the ethanol extracts of Auricularia auricula-judae depending on the solvent fractions and tumor cell lines. Further in vitro and in vivo studies will provide more information on the active compounds responsible for these activities and their potential as an anti-cancer remedy.

핑크팝 보리수 열매와 잎을 기능성 식품의 재료로 활용하기 위하여 항산화 및 총 페놀성 화합물 함량을 측정한 결과는 다음과 같다. DPPH 및 ABTS radical 소거활성을 측정한 결과 분획물의 농 도가 증가함에 따라 DPPH 및 ABTS radical 소거활성이 증가하는 경향을 보였으며, 특히 잎의 부 탄올 분획물에서 가장 높은 DPPH radical 소거활성을 보였다. 또한 ABTS 라디칼 소거활성도 열매 와 잎의 여러 분획물 중 잎의 부탄올 분획물에서 가장 높은 소거활성을 보였다. 환원력과 FRAP 활 성도 분획물의 농도가 증가함에 따라 환원력과 FRAP 활성이 증가하는 경향을 보였으며, 잎의 부탄 올 분획물에서 가장 높은 환원력과 FRAP 활성을 보였다. Linoleic acid를 이용한 자동산화 억제활 성을 실험한 결과 다른 분획물에 비하여 부탄올 분획물에서 가장 높은 과산화 억제활성을 나타내었 으며, 총 페놀성 화합물은 잎과 열매의 부탄올 분획물에서 각각 106.34 및 252.46 mg/g으로 가장 높은 함량을 보였다. 핑크팝 보리수 잎 부탄올 분획물의 항산화 활성은 페놀성 화합물에 의한 것이 며, 이는 자유 라디칼에 의해 유발된 산화적 스트레스를 효과적으로 방어할 수 있을 것이다.

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.

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.

The leaves and stems of Dendropanax morbiferus were separated from organic solvents with methanol. The organic solvent fractions were fractionated with dichloromethane, ethyl acetate and butanol according to the systematic fractionation method. Oxidation in the body induces aging, and antioxidant activity has attracted the attention of many people as a preventive component to suppress negative reactions in the body. To investigate the antioxidant activity of Dendropanax morbiferus were subjected to DPPH free radical assay. In addition, acetyl cholinesterase inhibitions were performed for Alzheimer’s disease as an aging neurological disease. As a result, it was confirmed that the antioxidant effect of DPPH was generally good in the antioxidant test. The ethyl acetate fractions of Dendropanax morbiferus stems and leaves were IC50=30 ㎍/㎖. Acetyl cholinesterase inhibition experiments were carried out at a concentration of 250 ㎍/㎖. Dendropanax morbiferus stems fractions showed dichloromethane fraction of 57.68%, which significantly inhibited the activity of acetyl cholinesterase.

This study was conducted to evaluate the cytotoxicity and α-amylase, α-glucosidase, pancreatic lipase inhibition in vitro by different solvent fractions from the roots of Codonopsis lanceolata. The values of IC50 against Calu-6 cell showed a high effect in n-hexane fraction (10.13 μg/mL) whereas DW fraction exhibited the weakest inhibition on cell viability, having an IC50 value of over 1,000 μg/mL. The values of IC50 against HCT-116 cell showed the highest activity in n-BuOH fraction (102.01 μg/mL), followed by n-hexane fraction (145.85 μg/mL), methylene chloride fraction (332.02 μg/mL), ethyl acetate fraction (462.93 μg/mL) and DW fracion (>1,000 μg/mL). α-Amylase inhibitory activity in methylene chloride fraction and ethyl acetate fraction was found to have a higher inhibitory effect with 24.5% and 25.6% than the other fractions. The highest α-glucosidase inhibitory activity was observed from the ethyl acetate fraction extract, while the extract of DW fraction showed the lowest level of inhibitory activity at given experiment concentration. The pancreatic lipase inhibitory activity of C. lanceolata was found to have a higher the effect in ethyl acetate fraction. Inhibition of lipase activity of the ethyl acetate fraction and n-hexane fraction showed a relatively high, while the extract of DW fraction showed the lowest level at given experiment concentration. These results suggested that the roots of C. lanceolata may assist in the potential biological activity on carbohydrate, lipid Inhibitory activity and anticancer activity.