본 연구에서는 생체재료인 콜라겐과 합성 단량체인 아크릴아마이드를 연속가교 하여, 하이드로젤 기반의 콜라겐 겔을 제조하였다. 아크릴아마이드의 함량 및 가교 정도에 따라, 1.5 kPa에서 3.0 kPa까지 다양한 강도 (E)를 갖는 콜라겐 겔을 제조할 수 있었다. 또한, 콜라겐 겔에 다공성 기공을 도입하고 진피세포를 내부에 담지하여, 겔 강도에 따른 세포 성장 및 거동을 확인하였다. 상대적으로 강도가 높은 겔에서 세포의 성장은 느렸지만 GAG 합성 및 분비는 활성화되는 것을 확인하였다. 콜라겐 겔의 기계적 물성에 따라 세포의 성장 및 활성이 영향을 받는 것을 알 수 있었으며, 이는 향후 인공피부 제조 및 응용, 나아가 다양한 조직공학 분야의 기반 기술로 활용 가능하리라 기대된다.

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.

Paeonia japonica is a perennial flowering plant used in traditional medicine therapy. The purpose of this study was to investigate the effect of water extract and solvent fractions obtained from P. japonica on anti-oxidative, anti-thrombin, anti-invasive and pro-apoptotic activities in YD-10B cells, human oral squamous carcinoma cell line. Water fraction revealed the highest extraction yield at 11.44% (w/w). Anti-oxidative activity was the highest in ethyl acetate fraction (85.13%). In the thrombin inhibitory activity test, ethyl fraction was the highest, with a value of 87.54%. Release and activation of MMP-2/pro-MMP-2 ratio in thrombin-treated YD-10B cells were significantly inhibited in the ethyl acetate fraction. At a concentration of 120 ㎍/㎖, water extract and solvent fractions of P. japonica inhibited cell proliferation in YD-10B cells except water fraction. Pro-apoptotic effect on human oral squamous carcinoma cell using the Bax/Bcl-2 ratio analysis was higher in water extract than other fractions. These findings suggest that the ethyl acetate fraction of P. japonica potentiates a promising antioxidant, anti-thrombin and anti-invasive agents.

As a part of an infrastructure project on medicinal herb-based remedies, we conducted a phytohemical investigation of the 100% MeOH extract from the aerial part of Boehmeria quelpaertense; our findings resulted in the isolation of flavonoids (1-2), isoquercitrin (1) and hyperoside (2). The identification and structural elucidation of these compounds were based on 1H-,13C-NMR, and LC ESI IT-TOF MS data. All the compounds isolated from this plant were reported for the first time. In this study, we examined the antioxidant activity of the 1 and 2 on the hydrogen peroxide (H2O2)-induced oxidative stress in a Rat Cardiomyoblast cell line (H9c2). The pretreatment of the flavonoids showed that it protects against H2O2-mediated cell death in the H9c2 cell line. Also, it decreases the intracellular reactive oxygen species (ROS) levels by the flavonoids in the H2O2-treated H9c2 cell line. These results showed that the 1 and 2 are a source of antioxidants. As a result, they might be helpful in preventing the progress of various oxidative stress mediated diseases, including myocardial infarction.

The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and ABTS+ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were 217.04±2.98, 156.72±3.90, and 182.88±3.02 mg TAE/g, respectively, while the electron donating abilities at 1,000 μg/mL of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The ABTS+ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at 50 μg/mL of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

In this study, the DPPH free radical scavenging activity, antimicrobial effects, growth inhibition and cytotoxicity of herb extracts were determined to screen alternative antibiotics. Hot water extracts of 10 species herbs (Origanum vulgare, Monarda didyma, Echinacea purpurea, Ocimum basilicum, Mentha piperita, Melissa officinalis, Thymus vulgaris, Stevia rebaudiana, Rosmarinus officinalis, Matricaria recutica) were used. DPPH free radical scavenging activity of all herb extracts was ranged from 31.4-49.9%, and significantly great activties were found at Rosmarinus officinalis, Origanum vulgare and Matricaria recutica (P<0.05). Hot water extracts of monarda didyma, origanum vulgare, thymus vulgaris and rosmarinus officinalis showed greater antimicrobial activities compared to others. Additionally, those four extracts represented relatively low cytotoxicity compared to others. As a result, it was found that Origanum vulgare and Rosmarinus officinalis which possessed great antioxidant and antimicrobial activity with less cytotoxicity. So these two herb extracts can be used as an alternative of antibiotics for organic farming.

Background: Hair loss related syndromes are increasing due to environmental pollution and stress. Hair care products are mainly prepared by mixing chemicals and natural extracts, such as those obtained from medicinal plants. The purpose of this study was to investigate the effects of 70% ethanol extracts from the flowers of Calendula officinalis, fruit body of Phellinus linteus, and the whole plant of Houttuynia cordata on the growth of CCD-986 cells, hair follicle dermal papilla cells (HFDPC), and 3T3-L1 cells. Methods and Results: All sample extracts at all concentrations, except for that from P. linteus fruit body at 500㎍/㎖, were cytotoxic to CCD-986 cells. However, none of the sample extracts were cytotoxic to HFDPC. The lipid differentiation of 3T3-L1 cells regulates hair regeneration via secretion of platelet derived growth factor. The 70% ethanol extract of H. cordata whole plant promoted hair growth. Adipogenesis rate significantly increased in a treatment concentration-dependent manner. Conclusions: These results suggest that 70% ethanol extracts of C. officinalis flower, P. linteus fruit body and H. cordata could be used for the development of hair care products.

Background: An imbalance in energy intake and expenditure can cause obesity, which is a major risk factor for chronic diseases such as heart disease, type 2 diabetes, insulin resistance, cancers and hyperlipidemia. Methods and Results: In this study, we evaluated the anti-obesity effects of a water extract from the young leaves of barley sprout (BS) in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice (HF). Lipid accumulation measurement indicates that BS markedly inhibited adipogenesis by reducing lipid droplet production in a dose-dependent manner. Furthermore, the mRNA expression of adipogenic transcription factors peroxisome proliferator-activated receptor-γ and fatty acid synthetase, CCAAT/enhancer binding protein-α and fatty acid binding protein 4 in 3T3-L1 cells was significantly inhibited by BS treatment. In an in vivo test, the BSadministered group of HFD-induced mice showed less body weight gain, and lower liver and epididymal white adipose tissue weights. The BS-treated mice showed decreased serum levels of leptin and lipids compared to untreated HFD mice and the levels of adiponectin and the HDL-cholesterol/total cholesterol ratio increased. These results indicate that BS inhibits body fat accumulation by reducing the mRNA expression of lipogenesis transcription factors and increasing serum adipokine concentration in in vitro and in vivo tests. Conclusions: BS reduced high fat diet-induced weight gain and had a positive effect on dyslipidemia.

피부에 가해지는 스트레스는 헤어조절 및 사이클에 직⋅간접적으로 중요한 영향을 미친다고 알려져 있 다. 특히, 모근세포는 스트레스에 의한 부신피질관련호르몬과 세포손상 및 사멸과 밀접한 관련이 있다고 보고되 고 있지만, 현재까지 실험적으로 입증된 사실은 매우 제한되어 있다. 보고에 의하면, 부신피질자극호르몬방출인 자가 증가되면 모근세포의 마이토콘드리아 활성을 저해하여 초기단계의 세포사멸을 가져올 수 있다고 임상학적 으로 보고된바가 있다. 특히 아토피 피부염으로 인한 스트레스는 부신피질자극호르몬방출인자와 부신피질관련 호르몬의 양을 증가시키며, 이는 모발의 outer epithelial sheath에 영향을 준다고 알려져 있으며, 이러한 스트 레스의 변화는 마이토콘드리아 손상을 초래하여 초기단계세포손상을 준다고 한다. 따라서 본 연구는 아토피피부 염스트레스가 피부의 모근세포에 주는 영향에 대하여 연구를 하였는데, 이에 대한 연구는 현재까지 전무한 실정 이다. 우리는 NC/Nga 마우스에 2,4-dinitrochlorobenzene (DNCB)로 아토피피부염을 유발 후, 피부 스트레스 생성에 의한 초기단계 세포손상을 스트레스관련 인자, 부신피질자극호르몬방출인자 및 그 관련 인자, annexin V 및 마이토콘드리아 반응을 이용하여 연구하였다. 그 결과, 아토피피부염에 의한 스트레스는 체내의 부신피질 자극호르몬방출인자 및 관련인자의 활성을 증가시킬 뿐 아니라, 모근세포에 영향을 주어 초기단계세포사멸을 초래하는 것으로 나타났다. 이는 아토피피부염관련 헤어손상을 일으킨다는 중요한 연구결과를 보고하는 바이며, 부신피질자극호르몬 조절관련 의약품 및 화장품 등과 같은 보조적 요법이 필요함을 제안한다.

8종의 잿빛곰팡이병 균주를 순무잎에 접종하여 병반의 크기를 확인한 결과 가장 강한 감염력을 보인 ‘포도-01’ 균주와 병반의 확산이 가장 적은 ‘오랜지’를 선발하였다. 순무잎이 저항성을 보인 ‘오랜지’균주를 처리한 잎이 감수성을 보인 ‘포도-01’균주를 처리한 잎보다 indole-3-ylmethyl glucosinolate (I3M-GLS) 함량이 무처리 보다 2.5배 이상 높았으나 ‘포도-01’ 균주를 처리한 잎에서는 무처리 보다 낮은 함량을 보였다. 균주의 메탄올 추출액과 물추출물을 식물배양세포에 처리한 결과 ‘오랜지’균 주의 추출물이 ‘포도-01’ 균주의 추출물보다 배양세포의 생장을 더 강하게 억제 한 것으로 나타났는데 ‘오랜지’ 균주의 메타놀 및 물 추출물 처리에서 배양세포의 활력은 각각 22.7% 및 16.5% 감소시키는 것으로 나타났다. 한편 ‘오랜지’균주 추출물을 처리 한 배양세포에서 I3M-GLS의 생합성이 ‘포도-01’ 균주 추출물 보다 현저히 높은 것으로 나타났다. 본 결과로 보아 식물체내에 생합성되는 I3M-GLS 함량은 잿빛곰팡이균에 대한 식물세포의 저항성과 밀접한 관계가 있는 것으로 판단된다.

Background: In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of the root of Aralia cordata var. continentalis (Kitagawa) Y. C. Chu (RAc-E70) against human colorectal cancer cells. Methods and Results: RAc-E70 suppressed the proliferation of the human colorectal cancer cell lines, HCT116 and SW480. Although RAc-E70 reduction cyclin D1 expression at the protein and mRNA levels, RAc-E70-induced reduction in cyclin D1 protein level occurred more dramatically than that of cyclin D1 mRNA. The RAc-E70-induced downregulation of cyclin D1 expression was attenuated in the presence of MG132. Additionally, RAc-E70 reduced HA-cyclin D1 levels in HCT116 cells transfected with HA-tagged wild type-cyclin D1 expression vector. RAc-E70-mediated cyclin D1 degradation was blocked in the presence of LiCl, a GSK3β inhibitorbut, but not PD98059, an ERK1/2 inhibitor and SB203580, a p38 inhibitor. Furthermore, RAc-E70 phosphorylated cyclin D1 at threonine-286 (T286), and LiCl-induced GSK3β inhibition reduced the RAc-E70-mediated phosphorylation of cyclin D1 at T286. Conclusions: Our results suggested that RAc-E70 may downregulate cyclin D1 expression as a potential anti-cancer target through GSK3β-dependent cyclin D1 degradation. Based on these findings, RAc-E70 maybe a potential candidate for the development of chemopreventive or therapeutic agents for human colorectal cancer.

Background: Cynaroside is a flavone, a flavonoid-like compound, known by different names (luteoloside and cinaroside). It is commonly found in Lonicera japonica Thunb., Chrysanthemum moriflium, and Angelica keiskei. The process of cell death has been classified as necrosis and apoptosis. Necrosis refers to unregulated cell death induced by a chemotherapeutic agent. Doxorubicin is an anthracycline anti-cancer drug used to treat acute leukemia, cancer, and lymphoma. However, it induces nephrotoxicity including tubular damage. Therefore, we investigated the protective effect of cynaroside against doxorubicin-induced necrosis in HK-2 cells. Methods and Results: To confirm the beneficial effect of cynaroside on doxorubicin-induced necrosis, HK-2 cells, a human proximal tubule epithelial cell line were treated with 10 μM doxorubicin and 80 μM cynaroside. Doxorubicin treatment resulted in increased DNA fragmentation, caspase-3 activity and mitochondria hyperactivation during cell necrosis. However, pretreatment with 80 μM cynaroside attenuated DNA fragmentation, caspase-3 activity and mitochondria hyperactivation induced by 10 μM doxorubicin in HK-2 cells. Conclusions: These results indicated that pretreatment with cynaroside ameliorated doxorubicin-induced necrosis in HK-2 cells. Therefore, cynaroside be used as a therapeutic agent for improving doxorubicin-induced nephrotoxicity. However, further studies are required to evaluated the toxicity of cynaroside treatment in animals and to determine its protective effect against doxorubicininduced nephrotoxicity in an animal model.

Background: Glycyrrhiza uralensis Radix (GR) is a crude drugs used in Asian countries that has been reported to prevent the progression of neurodegenerative diseases such as Alzheimer’s disease. The present study examined whether GR and its active compounds, glycyrrhizic acid (GA) and isoliquiritigenin (IL), exerted protective effects on H2O2-induced oxidative damage in C6 glial cells. Methods and Results: We exposed C6 glial cells to hydrogen peroxide (H2O2) for 24 h and investigated the cellular response to GR and its active compounds by evaluating cell viability, reactivie oxygen species (ROS) production, and apoptosis-related protein expression. GR successfully mitigated the reduced cell viability and ROS production induced by H2O2 in C6 glial cells, IL and GA significantly increased the cell viability and decreased ROS production. In addition, IL and GA down-regulated apoptotic Baxdependent caspase-3 activation, but each compound exerted different mechanisms, i.e., IL dose-dependently decreased ROS production and, GA up-regulated anti-apoptotic Bcl-2 expression. Conclusions: These results demonstrated that GR and its active components, IL and GA, exhibit potential for use as natural neurodegenerative agents for the modulation of apoptosis in C6 glial cells.

본 연구에서는 담쟁이덩굴 줄기 70% 에탄올 추출물과 발효균주 Lactobacillus pentosus를 이용하여 발효시킨 담쟁이덩굴 줄기 발효추출물에 대하여 항산화 및 세포보호 효과를 측정하였다. 1,1-Diphenyl-2-picrylhydrazyl (DPPH)를 이용한 자유라디칼 소거 활성(FSC50)은 담쟁이덩굴 줄기 추출물 및 발효추출물이 각각 42.3 및 34.5 μ g/mL로 발효 후의 라디칼 소거활성이 약 18.4% 더 높게 나타났다. Lumiol-의존성 화학발광법을 이용한 Fe3+-EDTA/H2O2계에서의 총 항산화능(OSC50) 평가에서도 담쟁이덩굴 줄기 추출물과 발효추출물은 각각 2.6 및 2.5 μ g/mL로 발효 후가 약 4.2% 정도 더 높은 총 항산화능을 나타냈다. 1O2로 유도된 적혈구 세포 손상에 있어서 추출물 및 발효추출물의 세포 보호 효과(τ50)는 50 μ g/mL에서 각각 126.4 및 173.0 min을 나타 내어 발효 후 세포 보호 효과가 약 34.0% 더 높게 나타났다. 발효추출물은 지용성 항산화제로 알려진 (+)-α -tocopherol (43.4 min)보다도 3.9배 높은 세포 보호 활성을 보여주었다. 사람 섬유아세포인 Hs68을 대상으로 elastase 저해 활성을 조사하였다. Elastase 저해 활성(IC50)은 담쟁이덩굴 줄기 추출물과 발효추출물에서 각각 873.6 및 687.8 μ g/mL로 발효 후에 elastase 저해 활성이 약 21.3% 더 높은 것으로 나타났다. 이상의 결과들은 담쟁이덩굴 줄기 발효추출물이 항산화 작용과 더불어 주름개선 효과를 가지는 천연 화장품 소재로써 응용 가능성 이 있음을 시사한다.

본 연구에서는 약콩(Glycine soja Siebold et Zucc.) 분획 추출물의 미백효능을 관찰하기 위해 B16F10 멜라노마 세포에서 TRP-1 (tyrosinase related protein-1), TRP-2 (tyrosinase related protein-2), 티로 시나제 발현을 평가하였다. 그 결과 약콩 분획 추출물 0.125, 0.25, 0.5, 2.0 mg/mL 농도에서 82% 이상의 높 은 세포생존율을 나타내었다. α-melanocyte stimulating hormone (α-MSH)을 처리한 B16F10 멜라노마 세포에 약콩의 EtOAc 분획 추출물을 처리한 결과 티로시나제 발현이 감소되었으며 TRP-1, TRP-2 단백질 발현이 감소하였다. 이러한 결과는 약콩 분획 추출물이 멜라닌생합성과 관련되는 단백질의 발현을 감소시켜 피 부 미백효능을 나타내는 것으로 기대할 수 있다.