Oxidative DNA damage negatively affects humans and the research is currently ongoing to find ways to reduce oxidative stress. Oxidative stress has been identified as a key factor in triggering various diseases. Thus, its alleviation is important for human health. Broussonetia kazinoki (B. kazinoki) has been used in traditional Korean medicine as a dermatological therapy to treat burns, pruritus, and acne. B. kazinoki is generally segregated into peeled root (PR), root bark (RB), peeled stem (PS), and stem bark (SB). To assess these components for their antioxidant activity and protection against DNA damage, their ethyl acetate fractions were examined by 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay. As a result of confirming the expression of factors involved in attenuating DNA damage, the protective effect of SB on oxidative stress suppressed the expression of p-p53 and γ-H2AX. Additionally, the levels of p53 and H2AX mRNA were significantly downregulated. In conclusion, these results indicated that the SB component of B. kazinoki had the potential to be used as an effective natural antioxidant compared to the other parts of the plant.

In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of branches from Taxillus yadoriki parasitic to Neolitsea sericea (TN-NS-B) against human lung cancer cells, A549. TY-NS-B dose-dependently suppressed the growth of A549 cells. TY-NS-B decreased β-catenin protein level, but not mRNA level in A549 cells. The downregulation of β-catenin protein level by TY-NS-B was attenuated in the presence of MG132. Although TY-NS-B phosphorylated β-catenin protein, the inhibition of GSK3β by LiCl did not blocked the reduction of β-catenin by TY-NS-B. In addition, TY-NS-B decreased β-catenin protein in A549 cells transfected with Flag-tagged wild type β- catenin or Flag-tagged S33/S37/T41 mutant β-catenin construct. Our results suggested that TN-NS-B may downregulate β- catenin protein level independent on GSK3β-induced β-catenin phosphorylation. Based on these findings, TY-NS-B may be a potential candidate for the development of chemopreventive or therapeutic agents for human lung cancer.

Background : Cudrania tricuspidata Bureau belonging to the mulberry family has attracted much attention as a nutrition ingredient many times as many as the mulberry tree. In this study, we examined the inhibitory effect of hot water extract of Cudrania tricuspidata Bureau fruits (CT-F) on inflammation and inflammatory diseases such as diabetes and gastiritis in cell-level experiments and animal models using mice. Methods and Results : The inhibitory effect of CT-F was investigated in LPS_stimulated RAW264.7 macrophages. Treatment with CT-F significantly inhibited the production of inflammatory mediators such as NO, IL-6 and TNF-a. The inhibitory effect of CT-F was shown to be related to inhibition of NF-kB activation in LPS-stimulated macrophages. In in vivo diabetic models using mice, treatment of CT-F lowered the level of blood glucose in alloxan-induce type-1 diabetic model as well as db/db type-2 mice model. CT-F also dramatically inhibited gastiritis induced by EtOH/HCl in mice. Conclusion : Hot water extract of Cudrania tricuspidata Bureau fruits was effective for inhibition of inflammation, diabetes and gastirits. Therefore, CT-F is thought to be a beneficial candidate for development of functional foods to prevent inflammation-associated diseases.

Background : Exposure to Ultraviolet B (UVB) causes oxidative stress, inflammation, pigmentation and severe skin damage. Astragalus membranaceus (AG) has been used as a traditional medicine and have been studied various physiological activities. During the roasting process, bioactive substances is change including antioxidant substances. The aim is study the antioxidant effects and reactive oxygen species (ROS) inhibitory effect of the roasted A. membranaceus (R-AG). on Human dermal fibroblast (HDF) cells. Methods and Results : To prepare of R-AG samples, roasting machine was used. AG and R-AG were extracted to water and 70% ethanol. AG samples were evaluated the antioxidant potential by measuring the 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2`-azino-bis 3-ethylbenzothiazoline-6-sulphonic acid (ABTS) scavenging activities. Additionally, total phenolic contents and total flavonoid contents was compared with antioxidant ingredients. AG and R-AGs were analyzed with HPLC determine the major compounds such as calycosin, mononetin and glycosides. The antioxidant activities of R-AG increased and changed in major compounds. In UVB exposed HDF cells, AGs did not affect cell viability and R-AG inhibited ROS more effectively than AG. Conclusion : From these results, R-AG can inhibit oxidative stress induced UVB in HDF cells.

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.

Pumpkin has long been used as traditional health materials in oriental medicine, pharmacy, medicine, and food industries in many countries. In this study, the water extract and two active components from tendril of young C. moschata Duch. were investigated on inflammation inhibitory activity. The water extract of young C. moschata Duch. showed high cell viability over 95% and it decreased the production of interlukin 6 (IL-6) and tumor necrosis factor (TNF-α) in the capacity of mouse bone marrow-derived macrophages (BMDM) upon lipopolysaccharide (LPS) stimulation. Also, isolated fraction (B4) suppressed the secretion of IL-6 and TNF-α. Among the domestically cultivated pumpkins, B4 fraction contained in the tendril part of them and it comprised of the order of tendril from Cucurbita pepo var. cylindrica, old of C. moschata Duch., and young of C. moschata Duch. These results suggest that water extracts of C. moschata Duch. and purified active compound, rutin, show anti-inflammation activity by suppression of the secretion of IL-6 and TNF-α. It can be applicable as pharmaceutical materials.

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.

Hericium erinaceus is considered a functional food and potential medicinal source. The present study was conducted to examine the potential antioxidant and anti-inflammatory activities of carried out with water and ethanol extracts of Hericium erinaceus grown on germinated green rice (HEGR-W and HEGR-E, respectively) and the water and ethanol extracts of germinated green rice (GR-W and GR-E, respectively) as potential medicinal resources or antioxidant and anti-inflammatory agents. Methods and Results: The total phenolic and flavonoid contents, DPPH, and ABTS activity, reducing power, DNA protective activity, cell viability, and NO production were investigated. The total phenolic and flavonoid contents were highest in HEGR-E (66.53 ± 2.40 ㎎·GAE/100 g and 82.12 ± 7.10 ㎎·CE/100 g respectively). HEGR-E exhibited high DPPH (44.70 ± 1.28%) and, ABTS (44.70 ± 1.28%) activity and reducing power (0.219). HEGR and GR extracts showed protective activity against DNA damage. The cytotoxicity of HEGR and GR in RAW264.7 cells and LPS-induced RAW264.7 cells was low. HEGR-E and GR-W exhibited anti-inflammatory effects through a 28% inhibition of NO production in LPS-induced RAW264.7 cells. Conclusions: These results suggested that the extracts of Hericium erinaceus grown on germinated green rice could be a potential medicinal material with natural antioxidant and NO inhibitory properties.

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.

In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.

In this study, we produced the recombinant lunasin peptide using E. coli and P. pastoris, and evaluated biological activity of the recombinant lunasin peptide. Lunasin peptide was produced from E. coli transfected with pPGEX-lunasin expression vector and P. pastoris GS115 transfected with pPIC-lunasin expression vector. These recombinant lunasin peptides were similar to the synthetic lunasin peptide in the identification by LC-ESI-MS. In addition, the recombinant lunasin peptide from E. coli and P. pastoris was bound in the chromatin, and inhibited histone acetylation and the activity of histone acetyltransferase. These findings suggest that the production of the lunasin peptide using E. coli and P. pastoris will be useful for industrial utilization of lunasin peptide.

밀착연접(tight junction, TJ)은 인접하는 표피 세포 사이를 서로 연결 및 접합하여 전해질과 수분의 이동을 조절할 뿐만 아니라 세포 내 신호를 전달하고 세포분열을 조절하는 등 다양한 기능을 갖고 있는 것으로 알려졌다. 또한 최근 연구에 따르면 TJ 관련 단백질들의 비정상적 발현은 암 발생 및 진행과 밀접한 관련이 있는 것으로 보고되었으며, TJ 구성 단백질의 발현 조절은 피부 장벽 강화 및 보습 조절과 연관된 것으로 알려졌다. 본 연구에서는 세포 장벽 조절을 통해 피부 보습 조절에 관여하는 새로운 화장품 소재를 발굴하기 위해 여러 가지 소재들에 대한 스크리닝을 수행하였다. 이 중 인공 감미료 소재로 널리 사용되는 스테비올 및 당 유도체(스 테비오사이드)의 미백 및 주름 개선 등의 효능에 대한 기존 보고에 따라, 이들에 의한 TJ 조절 메커니즘을 확인 하기 위해 다양한 세포 활성 기능 시험을 수행하였다. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt)를 이용한 실험을 통하여 스테 비올은 human keratinocyte cell line인 HaCaT 세포에 250 μ M 까지 독성을 나타내지 않음을 확인하였다. Quantitative real-time PCR을 이용한 TJ 관련 단백질들의 mRNA 발현 변화를 통하여 스테비올에 의한 TJ 조절 기능을 확인하였다. 그 결과, 스테비올은 TJ 관련 단백질 중 특이적으로 claudin 8을 대조군 대비 30% 수준까지 감소시키는 것을 관찰하였다. 또한, 세포이동에 의한 영향을 관찰한 결과 스테비올 처리에 의해 세포이 동이 현저히 저해되는 것을 확인하였다. 마지막으로 세포 장벽의 투과성 변화를 관찰하기 위해 표피세포 피부저 항(transepithelial electric resistance, TEER) 분석 결과 스테비올에 의한 세포투과성(cell permeability) 또 한 증가되는 것을 관찰하였다. 이에 반해, 스테비올 유도체(스테비오사이드, 리바우디오사이드)에서는 1000 μ M까지 세포 독성이 거의 나타나지 않을 뿐만 아니라 claudin 8 발현 억제 및 세포이동 저해현상도 관찰되지 않았다. 스테비올은 HaCaT 세포의 세포 독성, claudin 8 발현 억제, 그리고 세포 이동의 저해효과를 보이는 반면 스테비올 당 유도체인 스테비오사이드, 리바우디오사이드는 세포 독성 및 세포이동에 영향이 없는 것으로 나타난 본 연구 결과들은 스테비올 당 유도체가 향후 화장품 원료로써 스테비올보다 적합한 소재임을 시사한다

Background : Ischemic stroke is a common cause of adult disability and death worldwide. Excessive oxidative stress is an important pathogenic mechanism in ischemic stroke. Major reduction of endogenous antioxidative systems increases production of free radicals inducing peroxidation of lipid, protein, and nucleic acid. 1,3-Dipalmitoyl-2-oleoylglycerol (DPOG) is a triglyceride found in oils from various natural sources such as palm kernels, sunflower seeds and rice bran. We found DPOG as an active constituent of rice bran oil. In the present study, we investigated neuroprotective effect of DPOG derived from rice bran oil on excitotoxicity in cultured neurons and on ischemic brain injury in rats. Methods and Results : Transient focal ischemic brain damage was induced by 2 h middle cerebral artery occlusion followed by 24h reperfusion (MCAO/reperfusion) in rats. After MCAO/reperfusion, the infarct and edema volume of brain tissue was measured by 2,3,5-triphenyltetrazolium chloride (TTC) staining methods. Glutathione concentration and lipid peroxidation rate were measured in brain tissue. The expression levels of phosphorylated mitogen activated proteins kinases (MAPKs), inflammatory factors, and anti-apoptotic and pro-apoptotic proteins in brain tissue were detected by Western blot. Cerebral cortical neuronal cells were cultured in 15-days-old fetus. Cortical neurons were incubated with 1 mM N-methyl-D-aspartate (NMDA) for 14 h to produce excitotoxicity. Cell viability was measured by MTT assay. DPOG (1-5 mg/kg) significantly reduced MCAO/reperfusion-induced infarction and edema formation, neurological deficits, and brain cell death. Depletion of glutathione level and lipid peroxidation induced by MCAO/reperfusion were inhibited by administration of DPOG. The increase of phosphorylated MAPKs, inflammatory factors, and proapoptotic proteins and the decrease of antiapoptotic protein in ischemic brain were significantly inhibited by treatment with DPOG. DPOG (0.1-10 uM) inhibited 1 mM NMDA-induced neuronal cell death in cultured cortical neurons. Conclusion : From the above results, the present study provides an evidence that DPOG derived from rice bran oil might be effectively applied for the treatment of ischemic stroke.

Background : Alzheimer’s disease (AD) is a neurodegenerative disease characterized by progressive memory loss, cognitive impairment and personality defects accompanied by diffuse structural abnormalities in the brain. The major pathological hallmarks of AD include beta amyloid (Aß) protein deposition, presence of neurofibrillary tangles and neurodegeneration of cholinergic neurons. Aß, a 39-43 amino acid proteolytic fragment of amyloid precursor protein, is the major constituent of the senile plaques. Rice bran, the major byproduct of the rice milling industry, is the source of a high quality vegetable oil. Rice bran oil (RBO) has attracted much medicinal attention for its strong hypocholesterolemic properties because of its balanced fatty acid composition and high levels of antioxidant phytochemicals such as oryzanols, tocopherols and tocotrienols. The present study aims to investigate the protective effect of RBO against Aß (25-35)-induced neurotoxicity in in vitro and in vivo. Methods and Results : Memory impairment was produced by intracerebroventricular (i.c.v) microinjection of 15 nmol Aß (25-35) and measured by passive avoidance test in ICR mice. Glutathione concentration, lipid peroxidation rate and acetylcholine esterase activity were measured in mice brain. The expression levels of phosphorylated mitogen activated proteins kinases (MAPKs), inflammatory factors, and anti-apoptotic and pro-apoptotic proteins in mice brains were detected by Western blot. Cerebral cortical neuronal cells were cultured from 15-days-old fetus. Cortical neurons were incubated with 10 μM Aß (25-35) for 36 h. Cell viability was measured by MTT assay. Chronic treatments of RBO (0.1-1 ml/kg, 8 days, p.o.) protected against memory impairment induced by Aß (25-35). Depletion of glutathione level, lipid peroxidation and increased acetylcholine esterase activity by the treatment with Aß (25-35) were inhibited by administration of RBO. The increase of phosphorylated MAPKs, inflammatory factors, and proapoptotic proteins and the decrease of antiapoptotic protein in Aß (25-35)-administered mice brain were significantly inhibited by treatment with RBO. RBO (0.1-5ul/ml) inhibited 10μM Aß (25-35)-induced neuronal cell death in cultured cortical neurons. Conclusion : The present study suggests the role of RBO as a promising therapeutic for neurodegenerative diseases like AD and stroke.

Background : Recently, many studies to seek for medicinal herb compounds from natural products have been attempted capable of being free from harmful side effects and reducing the economic burden. This study was conducted to investigate whether Morus alba L.(MAL) water extracts has a biological safety, and an inhibitory potential against the mutagenicity induced by cigarette smoke condensates(CSC). Methods and Results : MAL was extracted with 70% ethanol and the yield of the extract was 35.1%. The 70% ethanol MAL extract was fractionated sequentially by diethylether, chloroform, dichloromethane, and water, respectively. Crude MAL 70% ethanol extract itself and its solvent fractions did not show any mutagenic effect up to 1 mg/plate toward Salmonella typhimurium TA 98 with or without the microsomal S-9 mixture of liver metabolic activations. On the other hand, the crude MAL extract showed an inhibitory activity against the mutagenicity of CSC with S-9 mixture. Diethyl ether layer among five solvent fractions showed the highest inhibitory activity at the same dose. The inhibitory activity of diethyl ether layer was also increased dose-dependently and the inhibitory rate was about 97.1% at the concentration of 1 mg/plate. Conclusion : In this study, we concluded that all of the crude MAL 70% ethanol extract and its solvent layers are potentially safe for mutagenicity and the diethyl ether layer among the crude MAL 70% ethanol extract has an inhibitory effect against the mutagenicity of CSC.

Antioxidant activity and inhibitory effect on oxidative DNA damage of ethyl acetate fractions extracted from Cone of Red Pine (Pinus densiflora) were investigated to find utilization of Cone, by-product of Red Pine, thrown out after berry shatter, as a new natural plant resource. Cone from P. densiflora was extracted with methanol (MeOH) and separated to petroleum ether, ethyl acetate and water fraction. Among them, ethyl acetate fraction was used. The antioxidant activity was conducted by the 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical, 2, 2'-Azino-bis (3-ethylbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS) radical scavenging assay, Fe 2+ chelating assay and reducing power assay. The inhibitory effect on oxidative DNA damage was determined by DNA cleavage assay using φX-174 RF I plasmid. The results of DPPH and ABTS radical scavenging activity at 200 ㎍/㎖ of extracts were 86.50% and 95.80% respectively, which were similar figures compared with L-ascorbic acid as control. Fe 2+ chelating activity was 77.96% and reducing power was 0.77 at 200 ㎍/㎖. Total phenolic component was 27.29±0.3 ㎎/g and Vitamin C content was 1.84±0.1 ㎎/g. Also ethyl acetate fraction from Cone has inhibitory effect, using φX-174 RF I plasmid on DNA cleavage assay. In conclusion, Cone, by-product of P. densiflora, showed high antioxidant activity and inhibitory effect on oxidative DNA damage. Therefore this study suggests Cone, useless by-product, can be developed as a new natural plant resource with lots of utilization such as an effective antioxidant, natural medicine, food, cosmetics and so on.